bluuebunny/crossref_metadata_2025 huggingface dataset:
Global City Networks and the Spread of Global Ideas
Authors: Sukarieh Mayssoun
Published: January 2024

This chapter considers the most important theories developed for talking about the global spread of dominant ideas in the context of global capitalist society. The chapter covers cultural imperialism; cultural and public diplomacy; policy diffusion, transfer, and mobility; and cultural globalization. It also highlights theories that offer valuable insights into the global spread of ideas and concepts such as globalization and policy diffusion. The chapter talks about the concept of cultural imperialism, which tends to endorse a crude, reductionist understanding of global power and domination. It highlights how concepts would benefit by being supplemented with a close attention to the spatiality and geography of ideas and knowledge.
Promises of an Afterlife
Authors: Herrlinger Page
Published: January 2024

This chapter describes the encounter between Joseph Stalin, the supreme atheist responsible for the deaths of millions, and “Brother Ioann” Churikov, an elderly but feisty man of God, as apocryphal. It points out how Stalin's visitation of Brother Ioann in prison offers a framework for characterizing the relationship between trezvenniki and the Soviet state. It also highlights Brother Ioann as one of the believers who suffered for their convictions under Stalin's rule and died in captivity in 1933. The chapter discusses the significance of living socialism for many believers, which meant adapting to the secular demands of Soviet life while finding ways to nurture their religious and spiritual selves. It details how the trezvenniki reimagined their lives in the context of official atheism during the 1930s–1960s, shaping their community, beliefs, and practices.
Epilogue
Authors: Herrlinger Page
Published: January 2024

This chapter explores how holy sobriety found fertile ground in post-Soviet Russia and the emergence of a new generation of trezvenniki as the Russian population struggled to survive the rapid increase in poverty, unemployment, and poor health conditions. It talks about Evgenii, a former IV drug addict that joined the sober community and observed a highly disciplined regimen of prayer and fasting in line with practices initiated by “Brother Ioann” Churikov. It also looks at Evgenii's belief that Brother Ioann was the Second Coming of Christ and his sentiment of the divinity of the elder Aleksei Ivanovich Sinnikov. The chapter reviews Brother Ioann's besedy and letters that were seen as a fixed voice of the living God ready to communicate with his followers. It notes the belief of trezvenniki in Vyritsa about living according to the laws of sobriety in order to bring both spiritual salvation and earthly salvation.
Settler Safe Zone or Colonial Staging Ground?
Authors: Blair James J. A.
Published: January 2024

This chapter draws on colonial letters, diaries, ordinances, and interviews with descendants of early settlers of the Falkland Islanders. It details how the settlement of the Falklands and the colonization of Patagonia became mutually constitutive. It also outlines the history of settlement and the movement of people between the islands and Patagonia. The chapter talks about the cattle frontier built on conditions of debt peonage, which resulted in the alienation and exile of South American gauchos after permanent British occupation. It addresses the legacy of a fraught missionary settlement in the Falklands and revisits the story of the Indigenous Yagán figure O'rundel'lico.
Imperial Diaspora
Authors: Blair James J. A.
Published: January 2024

This chapter analyzes social contradictions inherent to the colonial subjection of settlers and the Falkland Islanders' assertion of peoplehood. The chapter traces the local construction of a white ethnic community known as locally born kelpers. It describes how, since the 1982 war, Falklands society has become an imperial diasporic mixture of Islanders, British contractors, and military personnel as well as migrants. The chapter points out how citizenship remains contingent on degrees of imperial sovereignty and strict immigration laws and customs restrict access to the Falklands Way of Life. The chapter focuses on racialized power dynamics of authoritarian populism and self-determination that are fundamental for understanding imperial diaspora.
Regulation of lipid metabolism by APOE4 in intrahepatic cholangiocarcinoma via the enhancement of ABCA1 membrane expression
Authors: Qian Liqiang , Wang Gang , Li Bin , Su Haoyuan , Qin Lei
Published: January 2024

Intrahepatic cholangiocarcinoma (ICC) is a malignancy with a dismal prognosis, thus the discovery of promising diagnostic markers and treatment targets is still required. In this study, 1,852 differentially expressed genes (DEGs) were identified in the GSE45001 dataset for weighted gene co-expression network analysis (WGCNA), and the turquoise module was confirmed as the key module. Next, the subnetworks of the 1,009 genes in the turquoise module analyzed by MCODE, MCC, and BottleNeck algorithms identified nine overlapping genes (CAT, APOA1, APOC2, HSD17B4, EHHADH, APOA2, APOE4, ACOX1, AGXT), significantly associated with lipid metabolism pathways, such as peroxisome and cholesterol metabolism. Among them, APOE4 exhibited a potential tumor-suppressive role in ICC and high diagnostic value for ICC in both GSE45001 and GSE32879 datasets. In vitro experiments demonstrated Apolipoprotein E4 (APOE4) overexpression suppressed ICC cell proliferation, migration, and invasion, knockdown was the opposite trend. And in ICC modulated lipid metabolism, notably decreasing levels of TG, LDL-C, and HDL-C, while concurrently increasing the expressions of TC. Further, APOE4 also downregulated lipid metabolism-related genes, suggesting a key regulatory role in maintaining cellular homeostasis, and regulating the expression of the membrane protein ATP-binding cassette transporter A1 (ABCA1). These findings highlighted the coordinated regulation of lipid metabolism by APOE4 and ABCA1 in ICC progression, providing new insights into ICC mechanisms and potential therapeutic strategies.
Altered O-linked glycosylation in benign and malignant meningiomas
Authors: Talabnin Chutima , Trasaktaweesakul Thanawat , Jaturutthaweechot Pitchanun , Asavaritikrai Pundit , Kongnawakun Dusit , Silsirivanit Atit , Araki Norie , Talabnin Krajang
Published: January 2024

Background

Changes in protein glycosylation have been reported in various diseases, including cancer; however, the consequences of altered glycosylation in meningiomas remains undefined. We established two benign meningioma cell lines—SUT-MG12 and SUT-MG14, WHO grade I—and demonstrated the glycan and glycosyltransferase profiles of the mucin-type O-linked glycosylation in the primary benign meningioma cells compared with two malignant meningioma cell lines—HKBMM and IOMM-Lee, WHO grade III. Changes in O-linked glycosylation profiles in malignant meningiomas were proposed.

Methods

Primary culture technique, morphological analysis, and immunocytochemistry were used to establish and characterize two benign meningioma cell lines. The glycan profiles of the primary benign and malignant meningiomas cell lines were then analyzed using lectin cytochemistry. The gene expression of O-linked glycosyltransferases, mucins, sialyltransferases, and fucosyltransferases were analyzed in benign and malignant meningioma using the GEO database (GEO series GSE16581) and quantitative-PCR (qPCR).

Results

Lectin cytochemistry revealed that the terminal galactose (Gal) and N-acetyl galactosamine (GalNAc) were highly expressed in primary benign meningioma cells (WHO grade I) compared to malignant meningioma cell lines (WHO grade III). The expression profile of mucin types O-glycosyltransferases in meningiomas were observed through the GEO database and gene expression experiment in meningioma cell lines. In the GEO database, C1GALT1-specific chaperone (COSMC) and mucin 1 (MUC1) were significantly increased in malignant meningiomas (Grade II and III) compared with benign meningiomas (Grade I). Meanwhile, in the cell lines, Core 2 β1,6-N-acetylglucosaminyltransferase-2 (C2GNT2) was highly expressed in malignant meningiomas. We then investigated the complex mucin-type O-glycans structures by determination of sialyltransferases and fucosyltransferases. We found ST3 β-galactoside α-2,3-sialyltransferase 4 (ST3GAL4) was significantly decreased in the GEO database, while ST3GAL1, ST3GAL3, α1,3 fucosyltransferases 1 and 8 (FUT1 and FUT8) were highly expressed in malignant meningioma cell lines—(HKBMM)—compared to primary benign meningioma cells—(SUT-MG12 and SUT-MG14).

Conclusion

Our findings are the first to demonstrate the potential glycosylation changes in the O-linked glycans of malignant meningiomas compared with benign meningiomas, which may play an essential role in the progression, tumorigenesis, and malignancy of meningiomas.
Effect of the microstructure of 316L stainless steel processed by laser powder bed fusion on its wear performance
Authors:   , Barrionuevo Chiluiza Germán Omar
Published: January 2024

La fabricación aditiva de metal (AM) ha cambiado los paradigmas en el procesamiento de materiales. Sin embargo, al tratarse de un proceso de fabricación por capas y debido a la compleja interacción material-láser, la microestructura resultante difiere de las aleaciones homólogas convencionales, y su evolución depende del ciclo térmico, tanto del gradiente de temperatura como de la tasa de solidificación. Dado que las propiedades mecánicas dependen de la microestructura, las propiedades de una pieza fabricada por fusión selectiva láser (LPBF) están significativamente influenciadas por los parámetros de procesamiento y la estrategia de escaneo. Los componentes para aplicaciones de contacto cinemático requieren una evaluación del rendimiento de desgaste en el contexto de LPBF. La presente investigación se centra en el estudio del efecto de la potencia del láser, la velocidad de escaneo y el espaciamiento de la trama sobre la densidad, microdureza y desgaste del acero inoxidable 316L. Se utilizaron varias técnicas de caracterización para determinar cómo la microestructura afecta la resistencia al desgaste: microscopía óptica y electrónica, difracción de rayos X y espectrometría. Para la evaluación del desgaste se utilizó un tribómetro pin-on-disc y nanoindentación. Además, se realizó una simulación de elementos finitos para estudiar el comportamiento de la pileta fundida y determinar el gradiente térmico y la tasa de enfriamiento. Los resultados muestran que la velocidad es el único parámetro estadísticamente significativo que influye tanto en la densificación como en la microdureza. La muestra fabricada por LPBF muestra un aumento aproximado de 40% en la microdureza y una tasa de desgaste un 30% más baja que la muestra fabricada convencionalmente. La microestructura celular y columnar resultante de la ultrarrápida tasa de enfriamiento durante la solidificación permite una mayor deformación elástico-plástica y por lo tanto mejora la resistencia al desgaste. Finalmente, se concluye que la mejora en la resistencia al desgaste se debe a que la alta densidad de dislocaciones en el límite celular juega un papel clave en el efecto de fortalecimiento interfacial.
Impacts of Farm Business of School (FBS) Intervention on The Income of The Cocoa Farmers in Nigeria
Authors: Adegoroye Ademola , Oluwalade Temitope Abiodun , Adeyelu Adegoke Abidemi , Olorunfemi Oluwatosin Ayotomide , Mope Christianah
Published: January 2024

This study was designed to empirically investigate the impacts of Farm Business School (FBS) intervention on the income of cocoa farmers in some selected states in Nigeria. Primary data were collected through direct personal interviews, and with the use of a well-structured questionnaire from 300 sampled cocoa farmers. The data analytical techniques employed in this study include descriptive statistics, endogenous switching regression (ESR) and propensity score matching (PSM). The results from the descriptive statistics for participants and non-participants showed that most of the farmers were relatively old given life expectancy in Nigeria as 52 years. Participants and non-participants in the study area had a mean age of 54 years. The cocoa farming has been dominated by male farmers. The result of ESR was based on the average treatment effects of participants in FBS on income, and this shows that participation in FBS increases income significantly, and farmers that did not participate would have benefited significantly had they participated in FBS. PSM analysis indicated that participants in Farmers Business School and The PSM results imply that FBS training has increased the income of participants by 343,950.84 point. The results showed that participating in Farmers Business School leads to significant gains and impacts on income of cocoa farmers. Also, variables like level of education, farm size, amount of credit obtained, household size, and number of visits by extension agents have a significant impact on the level of participation. Therefore, research institutes and other agencies of government should improve upon their services of creating awareness for cocoa farmers to encourage the participation of more farmers in the training programme to have increased income. The combination of ESR and PSM analysis will also add value and contribute to the literature on the best approach to address impact analysis.
The Remkan Distribution and Its Applications
Authors: Uwaeme, O. R.  , Akpan, N. P.
Published: January 2024

Due to the ever growing demand for the development of new lifetime distributions to meet the goodness of fit demand of complex datasets, two-parameter distributions has been proposed in recent times. This study therefore aims to contribute to this demand. We propose a new two-parameter lifetime distribution known as the Remkan distribution. Important mathematical properties of the Remkan distribution such as the moments and other related measures, and moment generating function were derived and the model parameters estimated using the maximum likelihood estimate technique. Finally, the flexibility of the new Remkan distribution was illustrated using a real life dataset and the results showed that the new Remkan distribution was the best amongst other competing two parameter distributions.
Current State and Future Potential of Fisheries in the Mirsharai Coastal Areas of Chattogram for Enhancing Bangladesh’s Blue Economy
Authors: Mondal Md. Atiqul Islam , Siddiqui Abdullah Al Mamun , Rani Seema , Chowdhury Showmitra , Chowdhury Siraj Uddin Md. Babar
Published: January 2024

Aims: This study aims to comprehensively assess the fisheries resources in the Mirsharai coastal area of Chattogram district, focusing on their contribution to both local and national economies. The research seeks to identify challenges and vulnerabilities, particularly those arising from climate change, impacting pond owners, fish farmers, hatchery owners, and nursery owners in the region.

Study Design: The study adopts a cross-sectional design, employing field surveys and interviews to collect data on fish production, and challenges faced by stakeholders. The cross-sectional approach allows for a holistic understanding of the current state of fisheries resources and their interactions with climatic factors.

Place and Duration of Study: The research was conducted in Mirsharai, a coastal upazila of Chattogram district, Bangladesh. The study encompasses data collected during the January to December 2016, providing insights into the dynamics of fish production, and challenges faced by fisheries stakeholders in Mirsharai.

Methodology: Field surveys were conducted to gather quantitative data on fish production from both capture and culture fisheries. Additionally, interviews with pond owners, fish farmers, hatchery owners, and nursery owners were conducted to qualitatively assess the impact of challenges faced by fisheries stakeholders in Mirsharai on their operations and economic well-being. The combination of quantitative and qualitative methods ensures a comprehensive analysis.

Results: The average fish production in capture fisheries was determined to be 63 kg/ha, while culture fisheries, specifically pond aquaculture, yield an average of 1656 kg/ha. The results highlight the significant impact of challenges faced by fisheries stakeholders in Mirsharai on the local fisheries sector, revealing challenges related to market timing, post-larvae availability, and pricing that affect the economic sustainability of stakeholders.

Conclusion: The findings underscore the vulnerability of Mirsharai's fisheries to challenges faced by fisheries stakeholders in Mirsharai, emphasizing economic losses for fish farmers, hatchery owners, and nursery owners. The conclusion calls for urgent adaptive measures and climate-resilient strategies in the fisheries sector to ensure sustainability and viability in the face of evolving climatic conditions.

Implication: This study has implications for fisheries management and policy development, emphasizing the need for proactive measures to address climate-induced challenges and sustain the economic well-being of fisheries stakeholders in the Mirsharai coastal area.
Effects of Disease Complex of Penicellium and Root Knot Nematode on The growth and Development of Chilli (Capsicum annum L.) Crop
Authors: Samanta Sanjoy , Shelar Vaibhav Balasaheb , Banerjee Santanu , Doggalli Gangadhara , Hazarika Sanjay , Agrawal Smita , Singh Shivam , Shahni Yumkhaibam Sonia
Published: January 2024

The behaviour of fungal pathogens in a dormant or active state under different changes by infection of the host plant parasitic nematode was related to the function of fungi. Due to RKNs on the chilli in Meloidogyneinfected pot culture, Penicillium sp. demonstrated plant growth-promoting characteristics and nematocidal activities, according to the findings of the current study. This fungus can be used as a growth promoter in Treatments 2 and 3, where a mixture of Meloidogyne sp. and Penicelliumsp. showed higher growth rates and yellowing leaves in some pots, but no knots were found in the roots however, the current research may offer a theoretical basis for a growth promoter agent for better growth in Penicillium inoculated pots and In Nematode inoculated pots, Meloidogynesp can reduce plant growth. In both Meliodogynesp and Penicillium sp inoculated pots, it was demonstrated that decreasing nematode population increased the growth habits of chilies in terms of number of leaves Therefore, it has been demonstrated that Penicellium decreases nematocidal action while promoting growth parameters in chilli plants, and that using preventative measures can somewhat lessen insect infestation in control pots.
Prospecção de Patentes sobre o Cobre Refinado a Fogo
Authors: Costa Maria Ivonete Nunes , Lima Francisco Valdivino Rocha , Brandim Ayrton de Sá
Published: January 2024

O cobre possui diversas aplicações, sobretudo nos seguimentos elétricos e na construção civil, devido às excelentes propriedades, como a alta condutividade térmica e elétrica. A prospecção tecnológica é um método de mapear o progresso científico e a tecnologia que facilita a orientação de empresas e pesquisadores. Este artigo discute uma busca por patentes de cobre refinado a fogo. A metodologia utilizada foi a revisão sistemática, tomando como base de dados a plataforma do EPO, INPI e o Google Patents e busca nas bases de artigos Web of Science e Science Direct® is Elsevier®. Os resultados foram 35 patentes e 27 artigos relacionados, dos quais se observou que nos últimos anos o campo permanece no estágio embrionário, não havendo uma evolução significativa da tecnologia, com tendências na China, nos Estados Unidos, na Austrália, na Polônia, na França e na Alemanha. Na última década, notou-se maior difusão do tema na China, além de observar a necessidade de maiores investimentos na área, a fim incentivar a conexão entre empresas e universidades para estimular o desenvolvimento e a produção de inovações de técnicas de refino.
App “RevParkinson”: uma plataforma digital para a gestão do tratamento medicamentoso e acompanhamento da evolução da Doença de Parkinson
Authors: Francisco Viviane Cristina Cardoso , Mendonça Cláudio Márcio Campos de , Lima Rafael Pontes , Hage-Melim Lorane Izabel da Silva
Published: January 2024

A doença de Parkinson é uma condição relativamente comum que requer tratamento medicamentoso complexo para ser controlada. Isso pode gerar a diminuição da adesão ao tratamento devido aos efeitos adversos. Diante disso, foi idealizado um aplicativo móvel “RevParkinson”, que visa a aumentar a adesão ao tratamento e a melhorar a qualidade de vida dos pacientes. Este artigo descreve o processo de desenvolvimento do aplicativo, utilizando o modelo Canvas Business Model e Proposta de Valor, em um estudo exploratório e descritivo. A revisão bibliográfica foi discutida por uma equipe e subsidiou as informações documentadas por meio dos Canvas. O aplicativo foi submetido a testes técnicos e validação com usuários. Os resultados da validação demonstraram o potencial do aplicativo como instrumento capaz de aumentar a adesão à terapia medicamentosa em pessoas que vivem com a doença de Parkinson, pois atende aos requisitos propostos, podendo ter um impacto significativo na gestão da doença e na qualidade de vida dos pacientes, proporcionando um novo paradigma de cuidados em saúde digital.
BARRAGENS DE REJEITOS DE MINERAÇÃO: UMA PROPOSTA ALTERNATIVA NA PERSPECTIVA DA SUSTENTABILIDADE
Authors: Agra Filho Severino Soares
Published: January 2024

Este artigo pretende mostrar que os episódios de rompimentos de barragens destinadas ao depósito de rejeitos de mineração revelaram as deficiências e fragilidades do licenciamento ambiental estão relacionadas à flexibilização de excepcionalidade estabelecida no Código Florestal e na Política Nacional de Segurança de Barragens. As excepcionalidades previstas denotam uma grave inobservância, como também um flagrante conflito, com os objetivos e diretrizes preconizados nos dispositivos da Constituição Federal, da Política Nacional de Meio Ambiente e da Política Nacional de Recursos Hídricos, quais sejam a harmonização do desenvolvimento, em todas as suas dimensões, com a qualidade ambiental. A flexibilização induziu dificuldades na gestão sustentável das águas assim como na condução dos instrumentos necessários ao seu exercício sobretudo a aplicação do licenciamento ambiental, a autorização de supressão de vegetação em áreas protegidas e a outorga de intervenção nos sistemas hídricos. O presente trabalho se propõe à apreciação e discussão de uma proposta de resolução para os órgãos de gestão ambiental com medidas compatíveis e indutoras da gestão sustentável das águas. A proposta formulada foi concebida como uma forma de induzir uma dinâmica de pró-sustentabilidade ou de sustentabilidade progressiva preconizada na Agenda 21 Brasileira e de promover a melhoria contínua nas atividades de mineração.
ANÁLISE DAS ÁREAS AMBIENTALMENTE PROTEGIDAS NA BACIA DO RIO CAPIVARI EM CRUZ DAS ALMAS – BA A PARTIR DA UTILIZAÇÃO DOS DADOS DO CEFIR
Authors: De Santana Marins Daniela , Mendes Kruschewsky Lordelo Lidiane
Published: January 2024

O Cadastro Ambiental Rural (CAR) é um instrumento de gestão ambiental criado pela Lei nº 12.651/12 em nível federal ou, em âmbito estadual na Bahia, denominado Cadastro Estadual Florestal de Imóveis Rurais. Seu principal objetivo é compilar informações sobre propriedades rurais, como Reserva Legal e Áreas de Preservação Permanente, para controle, monitoramento e planejamento ambiental e econômico, visando combater o desmatamento. Essas informações são armazenadas em um banco de dados acessível ao público e podem subsidiar ações de monitoramento em áreas protegidas. A área da Bacia do Rio Capivari, localizada no município de Cruz das Almas (BA), foi selecionada como área de estudo devido à sua relevância para a segurança hídrica da região, além da preocupação com o atual cenário de uso e ocupação do solo da mesma. Diante disso, o presente estudo teve como objetivo geral analisar o atual cenário dos imóveis rurais localizados na bacia do Rio Capivari, com ênfase nas áreas ambientalmente protegidas, por meio do banco de dados do Cadastro Estadual Florestal de Imóveis Rurais (CEFIR), com o intuito de sinalizar a importância deste instrumento de regularização ambiental dentro da política de conservação ao meio ambiente. Para isso, foi realizada uma pesquisa bibliográfica e documental com abordagem qualitativa, visando um estudo detalhado dos marcos teóricos e legais; e a utilização da plataforma do Sistema Nacional de Cadastro Ambiental Rural (SICAR), para coleta dos dados e posterior confecção de mapas temáticos da área de estudo para discussão da conformidade com a legislação vigente. Por meio desse estudo foi possível detectar uma baixa adesão dos proprietários rurais, localizados na área de estudo, e informações com baixa confiabilidade por se tratar de um cadastro autodeclaratório, esse cenário torna-se um importante impulsionador na discussão estratégica sobre a atuação dos órgãos ambientais no processo de validação desse instrumento de gestão ambiental.
Suppression of Epidermal Growth Factor-Induced Phospholipase C Activation Associated With Actin Rearrangement in Rat Hepatocytes in Primary Culture
Authors: Nojiri Shunsuke , Hoek Jan
Published: June 2003

Hepatocytes maintained in primary culture for periods of 1 to 24 hours exhibited a rapid decline in epidermal growth factor (EGF)–induced activation of phospholipase C (PLC), as was evident in a loss of EGF–induced inositol 1,4,5–trisphosphate (IP3) formation and mobilization of Ca2+ from intracellular Ca2+ stores. The loss of PLC activation was not the result of a decrease in EGF receptor or phospholipase C–γ1 (PLCγ1) protein levels, nor the result of a loss of tyrosine phosphorylation of these proteins, but was associated with a decrease in EGF–induced translocation of PLCγ1 to the Triton–insoluble fraction, presumably reflecting binding to the actin cytoskeleton. Disruption of F–actin by treatment of cultured hepatocytes with cytochalasin D recovered the EGF–induced IP3 formation and Ca2+ mobilization to the same level and with the same dose–response relationship as was obtained in freshly isolated cells. Analysis of PLCγ1 colocalization with F–actin by confocal microscopy showed that PLCγ1 was mostly distributed diffusely in the cytosol, both in freshly plated cells and in cells in culture for 24 hours, despite marked differences in actin structures. EGF stimulation caused a modest redistribution of PLCγ1 and a detectable increase in colocalization with cortical actin structures in freshly plated cells or in cytochalasin D–treated cells, but in cells that had been maintained and spread in culture only a limited PLCγ1 relocation was detected to specific actin–structure associated with lamellipodia and membrane ruffles. We conclude that actin cytoskeletal structures can exert negative control over PLCγ1 activity in hepatocytes and the interaction of the enzyme with specific actin structures dissociates PLCγ1 tyrosine phosphorylation from activation of its enzymatic activity.
Silymarin in the Treatment of Patients With Primary Biliary Cirrhosis With A Suboptimal Response to Ursodeoxycholic Acid
Authors: Angulo Paul , Patel Tushar , Jorgensen Roberta A. , Therneau Terry M. , Lindor Keith D.
Published: June 2003

Ursodeoxycholic acid (UDCA) is a safe and effective medical therapy for most patients with primary biliary cirrhosis (PBC), but some patients show an incomplete response. Silymarin is a potent antioxidant with immunomodulatory and antifibrotic properties. The aim of this study was to evaluate the safety and assess the efficacy of silymarin in patients with PBC who had shown a suboptimal response to UDCA. Twenty–seven patients with PBC who had been on UDCA (13–15 mg/kg/day) therapy for 7 to 221 months and had shown a persistent elevation of alkaline phosphatase activity at least 2 times the upper limit of normal for more than 6 months were enrolled. Oral silymarin, 140 mg 3 times daily was given for 1 year, and patients continued on the same dosage of UDCA. No significant changes in serum alkaline phosphatase activity (897 ± 84 vs. 876 ± 95, P = .5), total bilirubin (0.9 ± 0.1 vs. 1 ± 0.1, P = .07), aspartate transaminase (AST) (58 ± 5 vs. 56 ± 6, P = .4), albumin (4.0 ± .06 vs. 4.1 ± .06, P = .4), or Mayo risk score (3.82 ± 0.2 vs. 3.88 ± 0.2, P = .4) were noted after 1 year of treatment with combination therapy. Transitory gastrointestinal adverse events occurred in 2 patients. In conclusion, although silymarin was well tolerated, this medication did not provide benefit to patients with PBC responding suboptimally to UDCA. The results of this pilot study would seem to discourage further controlled trials of silymarin in patients with PBC.
Intrahepatic Mrna Expression of Interferon–Inducible Antiviral Genes in Liver Diseases: Dsrna–Dependent Protein Kinase Overexpression and Rnase L Inhibitor Suppression in Chronic Hepatitis C
Authors: Yu Shin-Han , Nagayama Kazuyoshi , Enomoto Nobuyuki , Izumi Namiki , Marumo Fumiaki , Sato Chifumi
Published: June 2003

As a part of the defense mechanism of the host to viral infection, interferons induce the transcription of several genes. These interferon–inducible genes contribute to the eradication of the viruses. Whereas some studies suggested the participation of a dsRNA–dependent protein kinase in the host reaction to hepatitis C virus infection, the involvement of other interferon–inducible genes has not been evaluated. Furthermore, there has been no analysis on the expression profile of multiple interferon–inducible genes. The aim of this study was to clarify the hepatic mRNA expression profile of interferon–inducible genes with a special concern to chronic hepatitis C. A total of 76 liver biopsy samples (28 with chronic hepatitis C, 10 with chronic hepatitis B, 9 with alcoholic liver disease, 14 with autoimmune hepatitis, 10 with primary biliary cirrhosis, and 5 of normal liver) were enrolled. The expression of the following genes was quantified by competitive reverse transcription–polymerase chain reaction and was compared according to the etiology; dsRNA–dependent protein kinase (PKR), 2′,5′–oligoadenylate synthetase (2,5–AS), latent cellular endoribonuclease (RNase L), RNase L inhibitor, and MxA. As a result, PKR mRNA was significantly overexpressed in the liver of chronic hepatitis C compared with those of other etiologies (P= .0178), and it correlated significantly with serum alanine transaminase values (r = .51, P = .0054). Also, the expression of the RNase L inhibitor showed a significant reduction in chronic hepatitis C (P = .0184). The expressions of 2,5–AS, RNase L, and MxA were not different significantly irrespective to the etiology. In conclusion, hepatic overexpression of PKR and reduced expression of RNase L inhibitor seem to contribute to the anti–HCV mechanism characteristically.
Quantitative Dna Fragment Analysis for Detecting Low Amounts of Hepatitis B Virus Deletion Mutants in Highly Viremic Carriers
Authors: Schläger Folke , Schaefer Stephan , Metzler Markus , Gratzki Nils , Lampert Fritz , Gerlich Wolfram H. , Repp Reinald
Published: June 2003

Many variants of hepatitis B virus (HBV) with deletions in the viral genome have been identified. Some of these variants are indicator or even effector of a more severe course of hepatitis. These deletion mutants contribute a variable and sometimes very low proportion to the viral population. For early detection of small amounts of deletion mutants among a large number of wild–type genomes, we applied a new screening method designated quantitative fragment analysis (QFA). By QFA the whole viral genome can be scanned for the presence of deletions or insertions of ≥3 nucleotides representing more than 2% of the viral population. Using QFA we showed that an often described deletion of 8 nucleotides is packaged in viral capsids and not a polymerase chain reaction (PCR) artifact. QFA was applied to study the emergence of deletion mutants in a group of 18 pediatric patients who had been infected from a common source while being under multidrug cancer chemotherapy. All patients had developed a highly viremic asymptomatic HBV carrier state. In 3 of these patients 3 different kinds of HBV deletion mutants were found by QFA: 8 bp deletions within the core promoter, core gene deletions from 8 to 86 bp, and large deletions of up to 1,989 bp spanning the precore/core and the preS/S reading frames. PCR primers that specifically amplify deletion variants enabled the detection of additional patients harboring the investigated variant.
Chronic Ethanol Increases Adeno–Associated Viral Transgene Expression in Rat Liver Via Oxidant and NfκB–Dependent Mechanisms
Authors: Wheeler Michael D. , Kono Hiroshi , Rusyn Ivan , Arteel Gavin E. , McCarty Douglas , Samulski Richard Jude , Thurman Ronald G.
Published: June 2003

Recombinant adeno–associated virus (rAAV) transduction is limited in vivo, yet can be enhanced by hydroxyurea, ultraviolet–irradiation, or adenovirus coinfection, possibly via mechanisms involving stress in the host cell. Because chronic ethanol induces oxidative stress, it was hypothesized that chronic ethanol would increase rAAV transduction in vivo. To test this hypothesis, rAAV encoding β–galactosidase was given to Wistar rats that later received either ethanol diet or high–fat control diet via an enteral–feeding protocol for 3 weeks. Expression and activity of β–galactosidase in the liver were increased nearly 5–fold by ethanol. The increase in transgene expression was inhibited by antioxidant diphenylene iodonium (DPI), which is consistent with the hypothesis that ethanol causes an increase in rAAV transduction via oxidative stress. Ethanol increased DNA synthesis only slightly; however, it increased the nuclear transcription factor κB (NFκB) 4–fold, a phenomenon also sensitive to DPI. Moreover, a 6–fold increase in rAAV transgene expression was observed in an acute ischemia–reperfusion model of oxidative stress. Transgene expression was transiently increased 24 hours after ischemia–reperfusion 3 days and 3 weeks after rAAV infection. Further, adenoviral expression of superoxide dismutase or IκBα superrepressor inhibited rAAV transgene expression caused by ischemia–reperfusion. Therefore, it is concluded that ethanol increases rAAV transgene expression via mechanisms dependent on oxidative stress, and NFκB likely through enhancement of cytomegaloviral (CMV) promoter elements. Alcoholic liver disease is an attractive target for gene therapy because consumption of ethanol could theoretically increase expression of therapeutic genes (e.g., superoxide dismutase). Moreover, this study has important implications for rAAV gene therapy and potential enhancement and regulation of transgene expression in liver.
A Comprehensive Karyotypic Study on Human Hepatocellular Carcinoma by Spectral Karyotyping
Authors: Wong Nathalie , Lai Paul , Pang Elizabeth , Leung Thomas Wai-Tong , Lau Joseph Wan-Yee , Johnson Philip James
Published: June 2003

The current paucity of cytogenetic information on hepatocellular carcinoma (HCC) reflects the difficulties in culturing hepatocytes in vitro. Here, we report on the successful culture of 15 HCC cases. Chromosome aneuploidy ranging from a near–diploid to hyperhexaploid karyotype was found, but their complete karyotypic interpretations were hampered by the presence of many unidentifiable rearrangements. Spectral karyotyping (SKY) was used to elucidate structural changes in these HCC samples and 3 liver cancer cell lines (PLC/PRF/5, Hep3B, and HepG2). Frequent structural abnormalities were found on chromosomes 1 (13 of 15 cases; 3 of 3 cell lines), 8 (10 of 15 cases; 2 of 3 cell lines), 17 (9 of 15 cases; 3 of 3 cell lines), and 19 (9 of 15 cases; 1 of 3 cell lines). In particular, the chromosome regions 1p13–q21, 8p12–q21, 17p11–q12, 17q22, and 19p10–q13.1 were involved in multiple rearrangements. SKY analysis also suggested several previously undescribed breakpoints in HCC. These breakpoints, predominantly pericentromeric, clustered around the chromosome bands 2q33–q34, 3p13–q12, 4p14–q12, 5p10–q11, 7p12–q11, 10q10–q11, 11q10, 11q13–q21, 12q10–q13, 12q22–q23, 13q10–q14, 15q10, 16q10–q13, 18p11–q11, 20p11–q13.1, 21q10, and 22q10. When tumor sizes were compared, a significantly higher number of structural abnormalities was found in tumors larger than 4 cm (P = .007). Rearrangements such as t(1;8), t(1;11), t(1;19), and t(17;21) that were identified in both primary tumors and cell lines might represent markers that reflect proliferative advantages. Although SKY analysis did not indicate consistent translocations, it suggested nonrandom breakpoints, predominantly in the pericentromeric region, on a number of chromosomes. These breakpoint clusters may thus prove to be more important in the liver carcinogenesis and targets for further molecular investigations.
Early Detection of Viral Resistance by Determination of Hepatitis B Virus Polymerase Mutations in Patients Treated by Lamivudine for Chronic Hepatitis B
Authors: Si Ahmed Si Nafa , Tavan David , Pichoud Christian , Berby Françoise , Stuyver Lieven , Johnson Mark , Merle Philippe , Abidi Hassane , Trépo Christian , Zoulim Fabien
Published: June 2003

We have analyzed the molecular dynamics of emergence of drug–resistant strains in patients receiving lamivudine therapy for chronic hepatitis B. Twenty consecutive patients with lamivudine resistance were studied (13 hepatitis B e antigen [HBeAg]-positive patients and 7 HBe antibody [anti–HBe]-positive patients). Determination of viral genotype, precore mutants, and polymerase gene mutants (L528M, M552V, M552I) was performed using the research version of Lipa–HBV. Quantitative analysis of HBV DNA was performed using both branched DNA (bDNA) and polymerase chain reaction (PCR) assays. Polymerase mutants (genotypic resistance) were found in 16 of 20 patients. Genotypic resistance was detected earlier than the phenotypic resistance (P= .004). Quantitative PCR allowed detection of viral DNA throughout the entire study period in 16 of 20 patients. Analysis of pretreatment variables showed that high alanine transaminase (ALT) levels (>3 × the upper limit of normal [ULN]) was associated with a more rapid selection of drug–resistant mutants (P= .027) and a high hepatitis B virus (HBV) DNA level (>1,497 Meq/mL, bDNA) with a more rapid occurrence of phenotypic resistance (P= .04). At the time of viral breakthrough, the mean serum HBV–DNA values were not different from the pretreatment values (P= .37). ALT levels were higher in anti–HBe-positive patients compared with pretreatment values and to HBeAg–positive patients (P= .01). In 8 patients, antiviral therapy was modified after viral breakthrough, with the introduction of famciclovir and/or interferon alfa. Viral DNA became undetectable by bDNA in 3 patients who received interferon. Our results suggest that genotypic assays for polymerase mutant detection and quantitative determination of viremia with highly sensitive assay are warranted for an optimal monitoring of antiviral therapy of chronic hepatitis B.
Regulation of Ca2+ signaling in rat bile duct epithelia by inositol 1,4,5-trisphosphate receptor isoforms
Authors: Hirata Keiji , Dufour Jean-François , Shibao Kazunori , Knickelbein Roy , O’Neill Allison F. , Bode Hans-Peter , Cassio Doris , St-Pierre Marie V. , LaRusso Nicholas F. , Leite Fatima M. , Nathanson Michael H.
Published: June 2003

Cytosolic Ca2+ (Cai
2+) regulates secretion of bicarbonate and other ions in the cholangiocyte. In other cell types, this second messenger acts through Ca2+ waves, Ca2+ oscillations, and other subcellular Ca2+ signaling patterns, but little is known about the subcellular organization of Ca2+ signaling in cholangiocytes. Therefore, we examined Ca2+ signaling and the subcellular distribution of Ca2+ release channels in cholangiocytes and in a model cholangiocyte cell line. The expression and subcellular distribution of inositol 1,4,5-trisphosphate (InsP3) receptor (InsP3R) isoforms and the ryanodine receptor (RyR) were determined in cholangiocytes from normal rat liver and in the normal rat cholangiocyte (NRC) polarized bile duct cell line. Subcellular Ca2+ signaling in cholangiocytes was examined by confocal microscopy. All 3 InsP3R isoforms were expressed in cholangiocytes, whereas RyR was not expressed. The type III InsP3R was the most heavily expressed isoform at the protein level and was concentrated apically, whereas the type I and type II isoforms were expressed more uniformly. The type III InsP3R was expressed even more heavily in NRC cells but was concentrated apically in these cells as well. Adenosine triphosphate (ATP), which increases Ca2+ via InsP3 in cholangiocytes, induced Ca2+ oscillations in both cholangiocytes and NRC cells. Acetylcholine (ACh) induced apical-to-basal Ca2+ waves. In conclusion, Ca2+ signaling in cholangiocytes occurs as polarized Ca2+ waves that begin in the region of the type III InsP3R. Differential subcellular localization of InsP3R isoforms may be an important molecular mechanism for the formation of Ca2+ waves and oscillations in cholangiocytes. Because Cai
2+ is in part responsible for regulating ductular secretion, these findings also may have implications for the molecular basis of cholestatic disorders.
Expression, regulation, and function of αV integrins in hepatocellular carcinoma: An in vivo and in vitro study
Authors: Nejjari Mimoun , Hafdi Zakia , Gouysse Géraldine , Fiorentino Michelangelo , Béatrix Olivier , Dumortier Jérôme , Pourreyron Céline , Barozzi Chiara , D’Errico Antonia , Grigioni Walter F. , Scoazec Jean-Yves
Published: June 2003

The expression of αV integrins by neoplastic cells contributes to the promotion of local invasion and metastasis. The most characteristic extracellular ligands of αV integrins are vitronectin and fibronectin. Hepatocytes are the main source of vitronectin, and the capacity to synthesize and secrete vitronectin is usually retained in hepatocellular carcinoma. The aim of this study was to explore the expression, regulation, and functional role of αV integrins in hepatocellular carcinoma. We first analyzed the expression of αV integrins and their ligands fibronectin and vitronectin in 80 cases of hepatocellular carcinoma. αV integrin chain was detected in 44 cases and vitronectin in 50. Twenty-four of the 44 αV-positive tumors contained large amounts of vitronectin. These cases presented more frequently with adverse histoprognostic factors, including infiltrative growth pattern (62.5%), lack of capsule (71%), presence of capsular invasion (57%), and satellite nodules (50%). We then used HepG2 and Hep3B cell lines as in vitro models to study αV integrin regulation and function. HepG2 and Hep3B cells expressed αV integrin chain and used αVβ1 and αVβ5 for adhesion and migration on vitronectin. Tumor necrosis factor (TNF) α and transforming growth factor (TGF) β significantly increased the expression levels of αV integrins and stimulated the adhesion and migration of both HepG2 and Hep3B cell lines on vitronectin. The effects of growth factors on cell adhesion and migration were reproduced by incubation with conditioned medium from rat liver myofibroblasts. In conclusion, our results support the existence of an αV integrin/vitronectin connection in hepatocellular carcinoma and suggest that this connection may be an adverse prognostic factor.
Resistance surveillance in chronic hepatitis B patients treated with adefovir dipivoxil for up to 60 weeks
Authors: Yang Huiling , Westland Christopher E. , Delaney William E. , Heathcote Elizabeth J. , Ho Victoria , Fry John , Brosgart Carol , Gibbs Craig S. , Miller Michael D. , Xiong Shelly
Published: June 2003

Current therapies for chronic hepatitis B virus (HBV) infection do not provide adequate long-term control of viral replication in the majority of patients. Monotherapy with nucleoside analogs, such as lamivudine and famciclovir, is effective for short periods but results in the emergence of drug-resistant HBV in a substantial number of patients within 1 year of therapy. Adefovir dipivoxil (ADV) has demonstrated clinical activity against wild-type and lamivudine-resistant HBV, but it is unclear whether resistance mutations will emerge after long-term therapy with this drug. To determine whether extended treatment with ADV led to the emergence of drug-resistant populations of HBV, we analyzed virus isolated from patients currently enrolled in a long-term open-label study. The reverse transcriptase domain of HBV polymerase was amplified and sequenced from patients that had received a cumulative exposure of up to 60 weeks of ADV. During our analyses, several previously unreported amino acid substitutions were observed in the reverse transcriptase domain of HBV. Importantly, none of the observed mutations occurred in more than 1 patient, nor were they associated with an adefovir-resistant phenotype in vitro. Furthermore, none of the patients from whom these mutant viruses were isolated had evidence of virologic rebound. In conclusion, these results, although based on a limited number of patients, suggest that treatment with ADV does not lead to the emergence of resistant virus after up to 60 weeks of therapy.
Racial differences in effectiveness of α-fetoprotein for diagnosis of hepatocellular carcinoma in hepatitis C virus cirrhosis
Authors: Nguyen Mindie H. , Garcia Ruel T. , Simpson Peter W. , Wright Teresa L. , Keeffe Emmet B.
Published: June 2003

α-Fetoprotein (AFP) is frequently used as a diagnostic marker for hepatocellular carcinoma (HCC). Most available data concerning AFP come from studies of patients with chronic hepatitis B or other chronic liver diseases of mixed etiologies. Limited data concerning the diagnostic value of AFP for hepatitis C virus (HCV)-related HCC have to date come only from Asian and European studies, and results are conflicting. There may be significant differences in AFP levels depending on racial backgrounds and etiologies of primary liver disease. We conducted a multicenter, retrospective, case-control study of 163 HCC patients with HCV infection and 149 control patients with HCV-related cirrhosis. The positive likelihood ratios for AFP at 0 to 20, 21 to 50, 51 to 100, and 101 to 200 ng/mL were 0.46, 1.31, 1.15, and 6.90, respectively. No controls had AFP greater than 200 ng/mL. The sensitivity of AFP for the diagnosis of HCC in African Americans with HCV infection was lower than that of patients of all other ethnic groups combined (57.1% vs. 81.6% for AFP > 10 ng/mL, P = .02, and 42.9% vs. 66.0% for AFP > 20 ng/mL, P = .05). The area under the receiver operating characteristics curve was 0.81 for non-African Americans but only 0.56 for African Americans. In conclusion, AFP greater than 200 ng/mL can be used to confirm HCC in patients with HCV-related cirrhosis and a hepatic mass. However, AFP is insensitive for the diagnosis of HCC in African Americans.