bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of neutrophils in a mouse model of halothane-induced liver injury
Authors: You Qiang , Cheng Linling , Reilly Timothy P. , Wegmann Dale , Ju Cynthia
Published: November 2006

Drug-induced liver injury (DILI) is a major safety concern in drug development. Its prediction and prevention have been hindered by limited knowledge of the underlying mechanisms, in part the result of a lack of animal models. We developed a mouse model of halothane-induced liver injury and characterized the mechanisms accounting for tissue damage. Female and male Balb/c, DBA/1, and C57BL/6J mice were injected intraperitoneally with halothane. Serum levels of alanine aminotransferase and histology were evaluated to determine liver injury. Balb/c mice were found to be the most susceptible strain, followed by DBA/1, with no significant hepatotoxicity observed in C57BL/6J mice. Female Balb/c and DBA/1 mice developed more severe liver damage compared with their male counterparts. Bioactivation of halothane occurred similarly in all three strains based on detection of liver proteins adducted by the reactive metabolite. Mechanistic investigations revealed that hepatic message levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β); IL-6, and IL-8 were significantly higher in halothane-treated Balb/c mice compared to DBA/1 and C57BL/6J mice. Moreover, a higher number of neutrophils were recruited into the liver of Balb/c mice upon halothane treatment compared with DBA/1, with no obvious neutrophil infiltration detected in C57BL/6J mice. Neutrophil depletion experiments demonstrated a crucial role for these cells in the development of halothane-induced liver injury. The halothane-initiated hepatotoxicity and innate immune response-mediated escalation of tissue damage are consistent with events that occur in many cases of DILI.

In conclusion

, our model provides a platform for elucidating strain-based and gender-based susceptibility factors in DILI development.

Supplementary material for this article can be found on the HEPATOLOGY website
(http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html).
Fibrosis in genotype 3 chronic hepatitis C and nonalcoholic fatty liver disease: Role of insulin resistance and hepatic steatosis
Authors: Bugianesi Elisabetta , Marchesini Gulio , Gentilcore Elena , Cua Ian Homer Y. , Vanni Ester , Rizzetto Mario , George Jacob
Published: November 2006

Hepatic steatosis has been associated with fibrosis, but it is unknown whether the latter is independent of the etiology of fat infiltration. We analyzed the relationship between clinical characteristics, insulin resistance (HOMA-R) and histological parameters in 132 patients with “viral” steatosis caused by genotype 3 chronic hepatitis C (CHC-3) and 132 patients with “metabolic” steatosis caused by nonalcoholic fatty liver disease (NAFLD), matched by age, BMI, and degree of liver fat accumulation. Tests of liver function were comparable in the two study populations. The prevalence of features of insulin resistance was higher in NAFLD, as was HOMA-R (

P

= .008). Logistic regression analysis confirmed that steatosis was associated with a high viral load and low serum cholesterol in CHC-3, and with high aminotransferase, glucose, ferritin and hypertriglyceridemia in NAFLD. At univariate analysis, advanced fibrosis was associated with steatosis in NAFLD, but not in CHC-3. Other parameters related to fibrosis severity were HOMA-R and a low platelet count in CHC-3, and high aminotransferases, HOMA-R, ferritin and low HDL-cholesterol in NAFLD. On multivariate analysis, only low platelet count (OR = 0.78; 95% CI, 0.67-0.92) and HOMA-R (OR = 2.98; 1.13-7.89) were independent predictors of advanced fibrosis in CHC-3. In NAFLD, severe fibrosis was predicted by fat grading (OR = 3.03; 1.41-6.53), ferritin (OR = 1.13; 1.03-1.25) and HOMA-R (OR = 1.16; 1.02-1.31).

In conclusion

, insulin resistance is an independent predictor of advanced fibrosis in both NAFLD and CHC-3, but the extent of steatosis contributes to advanced disease only in NAFLD. Virus-induced hepatic steatosis as seen in CHC-3 does not contribute significantly to liver fibrosis.
Portopulmonary hypertension: Results from a 10-year screening algorithm
Authors: Krowka Michael J. , Swanson Karen L. , Frantz Robert P. , McGoon Michael D. , Wiesner Russell H.
Published: November 2006

Portopulmonary hypertension (POPH) is the elevation of pulmonary artery pressure due to increased resistance to pulmonary blood flow in the setting of portal hypertension. Increased mortality has occurred with attempted liver transplantation in such patients and thus, screening for POPH is advised. We examined the relationship between screening echocardiography and right heart catheterization determinations of pressure, flow, volume, and resistance. A prospective, echocardiography–catheterization algorithm was followed from 1996 to 2005. Consecutive transplantation candidates underwent Doppler echocardiography to determine right ventricular systolic pressure (RVSP). Of 1,235 patients, 101 with RVSP >50 mm Hg underwent catheterization to measure mean pulmonary artery pressure (MPAP), flow via cardiac output (CO), central volume via pulmonary artery occlusion pressure (PAOP), and resistance via calculated pulmonary vascular resistance (PVR). Bland-Altman analysis suggested marked discordance between echocardiography-derived RVSP and catheterization results. All-cause pulmonary hypertension (MPAP >25 mm Hg) was documented in 90/101 (90%) patients. Using current pressure and resistance diagnostic guidelines (MPAP >25 mm Hg, PVR ≥240 dynes/s/cm−5), POPH was documented in 66/101 (65%) patients. Elevated MPAP was due to increased CO and/or PAOP in 35/101 (35%) patients with normal resistance (PVR <240 dynes/s/cm−5). The transpulmonary gradient (MPAP–PAOP) further characterized POPH in the presence of increased volume. Model for end stage liver disease (MELD) scores correlated poorly with MPAP and PVR.

In conclusion

, right heart catheterization is necessary to confirm POPH and frequently identifies other reasons for pulmonary hypertension (

e.g.

, high flow and increased central volume) in liver transplantation candidates. Severity of POPH correlates poorly with MELD scores.
The protein kinase IKKε can inhibit HCV expression independently of IFN and its own expression is downregulated in HCV-infected livers
Authors: Vilasco Myriam , Larrea Esther , Vitour Damien , Dabo Stephanie , Breiman Adrien , Regnault Béatrice , Riezu Jose-Ignacio , Eid Pierre , Prieto Jesus , Meurs Eliane F.
Published: November 2006

During a viral infection, binding of viral double-stranded RNAs (dsRNAs) to the cytosolic RNA helicase RIG-1 leads to recruitment of the mitochondria-associated Cardif protein, involved in activation of the IRF3-phosphorylating IKKε/TBK1 kinases, interferon (IFN) induction, and development of the innate immune response. The hepatitis C virus (HCV) NS3/4A protease cleaves Cardif and abrogates both IKKε/TBK1 activation and IFN induction. By using an HCV replicon model, we previously showed that ectopic overexpression of IKKε can inhibit HCV expression. Here, analysis of the IKKε transcriptome profile in these HCV replicon cells showed induction of several genes associated with the antiviral action of IFN. Interestingly, IKKε still inhibits HCV expression in the presence of neutralizing antibodies to IFN receptors or in the presence of a dominant negative STAT1α mutant. This suggests that good IKKε expression levels are important for rapid activation of the cellular antiviral response in HCV-infected cells, in addition to provoking IFN induction. To determine the physiological importance of IKKε in HCV infection, we then analyzed its expression levels in liver biopsy specimens from HCV-infected patients. This analysis also included genes of the IFN induction pathway (RIG-I, MDA5, LGP2, Cardif, TBK1), and three IKKε-induced genes (IFN-β, CCL3, and ISG15). The results show significant inhibition of expression of IKKε and of the RNA helicases RIG-I/MDA5/LGP2 in the HCV-infected patients, whereas expression of TBK1 and Cardif was not significantly altered.

In conclusion

, given the antiviral potential of IKKε and of the RNA helicases, these

in vivo

data strongly support an important role for these genes in the control of HCV infection.

Supplementary material for this article can be found on the HEPATOLOGY website
(http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html).
Engineered measles virus as a novel oncolytic viral therapy system for hepatocellular carcinoma
Authors: Blechacz Boris , Splinter Patrick L. , Greiner Suzanne , Myers Rae , Peng Kah-Whye , Federspiel Mark J. , Russell Stephen J. , LaRusso Nicholas F.
Published: November 2006

The oncolytic measles virus Edmonston strain (MV-Edm), a nonpathogenic virus targeting cells expressing abundant CD46, selectively destroys neoplastic tissue. Clinical development of MV-Edm would benefit from noninvasive monitoring strategies to determine the speed and extent of the spread of the virus in treated patients and the location of virus-infected cells. We evaluated recombinant MV-Edm expressing carcinoembryonic antigen (CEA) or the human sodium iodide symporter (hNIS) for oncolytic potential in hepatocellular carcinoma (HCC) and efficiency in tracking viruses

in vivo

by noninvasive monitoring. CD46 expression in human HCC and primary hepatocytes was assessed by flow cytometry and immunohistochemistry. Infectivity, syncytium formation, and cytotoxicity of recombinant MV-Edm in HCC cell lines were evaluated by fluorescence microscopy, crystal violet staining, and the MTS assay. Transgene expression in HCC cell lines after infection with recombinant MV-Edm

in vitro

and

in vivo

was assessed by CEA concentration, 125I-uptake, and 123I-imaging studies. Toxicology studies were performed in IfnarKO×CD46 transgenic mice. The CD46 receptor was highly expressed in HCC compared to nonmalignant hepatic tissue. Recombinant MV-Edm efficiently infected HCC cell lines, resulting in extensive syncytium formation followed by cell death. Transduction of HCC cell lines and subcutaneous HCC xenografts with recombinant MV-Edm resulted in high-level expression of transgenes

in vitro

and

in vivo

. MV-Edm was nontoxic in susceptible mice. Intratumoral and intravenous therapy with recombinant MV-Edm resulted in inhibition of tumor growth and prolongation of survival with complete tumor regression in up to one third of animals.

In conclusion

, engineered MV-Edm may be a potent and novel cancer gene therapy system for HCC. MV-Edm expressing CEA or hNIS elicited oncolytic effects in human HCC cell lines

in vitro

and

in vivo,

enabling the spread of the virus to be monitored in a noninvasive manner.
Intrahepatic virus-specific IL-10-producing CD8 T cells prevent liver damage during chronic hepatitis C virus infection
Authors: Abel Michal , Sène Damien , Pol Stanislas , Bourlière Marc , Poynard Thierry , Charlotte Frédéric , Cacoub Patrice , Caillat-Zucman Sophie
Published: November 2006

CD8 T cell killing of hepatitis C virus (HCV)-infected hepatocytes is thought to contribute to liver damage during chronic HCV infection, whereas the participation of HCV-nonspecific immune cells is unclear. To visualize the spatial relationship of HCV-specific CD8 T cells with parenchymal target cells, and to examine their local functional activity in relation to hepatocellular necrosis and fibrosis, we used HLA tetramers and confocal microscopy in biopsies from 23 HLA-A2 or HLA-B7 patients with chronic HCV infection. Intrahepatic tetramer+ (HCV-specific) CD8 T cells protected from hepatic necroinflammatory disease activity, independently of age, gender, viral load, and viral genotype. Indeed, tetramer+ cells were scattered in the liver within regions of weak fibrosis (low laminin expression) and low hepatocellular apoptosis (TUNEL method), and expressed IL-10 but not IFNγ. By contrast, tetramer-negative CD8 T cells were associated with active necroinflammatory liver disease, colocalized with strong laminin expression and hepatocellular apoptosis, and expressed more frequently IFNγ than IL-10. Overall, liver regions harboring HCV-specific CD8 T cells tended to be healthier than areas containing only inflammatory cells of undefined specificity.

In conclusion

, HCV-specific IL-10-producing CD8 T cells, although not cytotoxic and unable to control viral replication, can attenuate hepatocellular necrosis, liver fibrosis, and inflammation mediated by bystander T cells, and may thus represent antigen-induced regulatory CD8 T cells. Therapeutic modulation of the intrahepatic balance between specific and bystander CD8 T cells might be beneficial in patients with chronic hepatitis C.

Supplementary material for this article can be found on the HEPATOLOGY website
(http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html).
Surprises from the crystal structure of the hepatitis C virus NS2-3 protease
Authors: Gouttenoire Jerome , Moradpour Darius , Penin François
Published: November 2006

Hepatitis C virus is a major global health problem affecting an estimated 170 million people worldwide. Chronic infection is common and can lead to cirrhosis and liver cancer. There is no vaccine available and current therapies have met with limited success. The viral RNA genome encodes a polyprotein that includes 2 proteases essential for virus replication. The NS2-3 protease mediates a single cleavage at the NS2/NS3 junction, whereas the NS3-4A protease cleaves at 4 downstream sites in the polyprotein. NS3-4A is characterized as a serine protease with a chymotrypsin-like fold, but the enzymatic mechanism of the NS2-3 protease remains unresolved. Here, we report the crystal structure of the catalytic domain of the NS2-3 protease at 2.3 Å resolution. The structure reveals a dimeric cysteine protease with 2 composite active sites. For each active site, the catalytic histidine and glutamate residues are contributed by one monomer, and the nucleophilic cysteine by the other. The carboxy-terminal residues remain coordinated in the 2 active sites, predicting an inactive postcleavage form. Proteolysis through formation of a composite active site occurs in the context of the viral polyprotein expressed in mammalian cells. These features offer unexpected insights into polyprotein processing by hepatitis C virus and new opportunities for antiviral drug design.
Genomics and complex liver disease: Challenges and opportunities
Authors: Juran Brian D. , Lazaridis Konstantinos N.
Published: November 2006

The concept of genetic susceptibility in the contribution to human disease is not new. What is new is the emerging ability of the field of genomics to detect, assess, and interpret genetic variation in the study of susceptibility to development of disease. Deciphering the human genome sequence and the publication of the human haplotype map are key elements of this effort. However, we are only beginning to understand the contribution of genetic predisposition to complex liver disease through its interaction with environmental risk factors. In the coming decade, we anticipate the development of human studies to better dissect the genotype/phenotype relationship of complex liver diseases. This endeavor will require large, well-phenotyped patient populations of each disease of interest and proper study designs aimed at answering important questions of hepatic disease prognosis, pathogenesis, and treatment. Teamwork between patients, physicians, and genomics scientists can ensure that this opportunity leads to important biological discoveries and improved treatment of complex disease.
Insiders and outsiders: Levels of collaboration in research partnerships across resource divides
Authors: TOMLINSON MARK , SWARTZ LESLIE , LANDMAN MIREILLE
Published: November 2006

With increasing recognition of the health and psychosocial challenges in developing countries, more research in these countries is essential. Research collaborations between wealthier and less wealthy countries are, however, complex. A partnership model has been proposed which emphasizes the importance of mutually respectful and beneficial relationships in international research. Using an example of a mother‐infant intervention project in South Africa we describe the elements of an equitable partnership model of research. An issue which has been overlooked, however, is the reality of power differentials within the poorer countries themselves. We show that there are many intersecting levels of relationships within intercountry research, and suggest that power dynamics within countries deserves equal attention as international issues. Based on our experiences, we present some tentative guidelines for international research which builds long‐term capacity and recognizes the importance of multiple levels of analysis and interaction.
Culturally sensitive assessment of attachment in children aged 18–40 months in a South African township
Authors: MINDE KLAUS , MINDE REGINA , VOGEL WENDY
Published: November 2006

The objective of our work is to study the possible relevance nonwestern cultural traditions have on the concordance of attachment patterns assessed in mothers and their young children. The attachment of 46 toddlers and their mothers, living in a black township in Johannesburg, South Africa, was assessed using scores derived from mother‐child observations in the home (Attachment Q‐Sort) and an interview (Working Model of the Child Interview). Mothers also had a semistructured psychiatric interview. Agreement between home observations and interview ratings was 29% for secure and 71% for insecure attachment when U.S.‐developed scoring criteria for the interview were used. Agreement increased to 81% for secure and 67% for insecure attachment when the same protocols were rescored, using a culturally modified scoring system, developed by local cultural experts. This study suggests that verbal representations of attachment patterns are more influenced by cultural traditions than are actual parent‐child interactions.
Infants and young children living in youth‐headed households in Rwanda: Implications of emerging data
Authors: BORIS NEIL W. , THURMAN TONYA R. , SNIDER LESLIE , SPENCER ERIN , BROWN LISANNE
Published: November 2006

The HIV/AIDS epidemic has already produced millions of orphans in Africa and the peak of the orphan epidemic has not yet been reached. One emerging social trend associated with premature parental death is the formation of households headed by youth aged 13–24. In Rwanda, specific sociocultural factors have supported the formation of youth‐headed households (YHH) in the wake of both the 1994 genocide and the ongoing HIV/AIDS epidemic. There is no published data on the health and socioemotional functioning of children under 5 living in YHH. Survey data from 692 YHH in one region of Rwanda yielded a subsample of 89 homes which contained one or more children under 5 (n=104). These data reveal that a majority of young children living in YHH's is in fair or poor health. Heads of household themselves report high levels of depressive symptoms and social isolation; reports of emotional distress among the youngest children in the homes are associated with reports of higher symptomatology among heads of household. Findings are interpreted in light of the sociocultural context of Rwanda and the implications of the data for intervention are considered.
Ecological approach to management of open drains
Authors: Lamsodis Romanas , Morkūnas Vytautas , Poškus Vaclovas , Povilaitis Arvydas
Published: November 2006

Most natural streams were made into drains when they were converted into recipients of subsurface tile systems. During the process the variety of biotic and abiotic conditions within and around the streams was totally destroyed, and the new watercourses (open drain recipients) were deprived of the greater part of the ecological importance that the former streams had possessed in the landscape. The data collected over the last decade in Lithuania about drain bed deformations, sediment accumulation, vegetation cover successions, intentional afforestation of slopes, beaver expansion and succeeding water quality changes, revealed some self‐perpetuating natural alterations which raise the ecological significance of open drains as refuges and corridors in the landscape for vegetation (the succession of herbage and appearance of woody vegetation on slopes), and the barriers protecting surface runoff from diffuse agricultural pollution (beaver impoundments). On the other hand, these alterations are useful in terms of maintenance of the functioning of those drains as recipients because they result in steadier (roughly parabola‐shaped) cross‐section profiles, less threat of bed silting up and no need for the mowing of slopes (because of the growth of woody vegetation). This paper presents and discusses the study results highlighting the above‐mentioned bidirectional effect of self‐renaturalization, and considers the preconditions and possible methods of introduction for the ecologically acceptable management of open drains. Copyright © 2006 John Wiley & Sons, Ltd.
Efficiency of old tile drain systems in soils with high clay content: differences in the trench backfill zone versus the zone midway between trenches
Authors: Messing I. , Wesström I.
Published: November 2006

This study was carried out in order to check the efficiency in water transport capacity in the trench backfill zone (IT) in 2–45‐year‐old tile drains, and the differences between IT and the zone midway between trenches (BT) in field‐saturated steady‐state infiltration rate (Ifs) and saturated hydraulic conductivity (Ks). At seven sites with high clay content, Ifs was recorded in situ in large rings (1 m diameter) and at the same time small cores (7.2 cm in diameter) were sampled to measure Ks in the laboratory. There were large differences in drainage efficiency between the ‘disturbed’ trench backfill zone and the ‘undisturbed’ zone midway between trench lines. Ifs and Ks values were in almost all cases larger in IT than in BT. Differences between the plough sole and subsoil, for both Ifs and Ks, were more evident in BT than in IT. The positive effect above the drainpipes, indicating that macropore connectivity was superior in IT, was evident for as long as 45 years after the installation of drainpipes. In BT, Ifs and Ks ranged within approximately equal values. In IT, however, Ifs ranged at larger values than Ks. In modelling these kinds of soils, it may be considered whether water transport during heavy rain events should be characterized by a quick onset of water flow in drainpipes caused by the more permeable soil in the backfill trench, and a subsequent slower and more continuous transport of water from the zone between trenches, and in what way the drain water quality is influenced by this. Copyright © 2006 John Wiley & Sons, Ltd.
Integrin α5 controls osteoblastic proliferation and differentiation responses to titanium substrates presenting different roughness characteristics in a roughness independent manner
Authors: Keselowsky B.G. , Wang L. , Schwartz Z. , Garcia A.J. , Boyan B.D.
Published: November 2006

Integrin α5β1 regulates osteoblast proliferation and differentiation on smooth synthetic surfaces presenting different chemistries, but it is not known whether this integrin controls osteoblast behavior on surfaces that have micron‐scale rough topographies. We cultured MG63 human osteoblast‐like cells on titanium substrates with three different roughness characteristics: chemically polished (PT), grit blasted and acid etched with a complex topography consisting of 20–100 μm craters and 0.5–2 μm micropits (SLA), and plasma‐sprayed Ti with irregular projections (TPS). Cells spread well on PT but displayed a smaller footprint on SLA or TPS. Nuclei were larger on PT as well. α5β1 binding and FAK phosphorylation were greater on the rougher surfaces (TPS > SLA > PT). Antibodies against the α5β1 binding site on fibronectin had no effect on cell number at 3 days, but [3H]‐thymidine incorporation was increased, suggesting that binding to fibronectin was necessary for cell cycle regulation. Antibodies to the α5 subunit reduced cell number at 3 days on PT and TPS and reduced DNA synthesis on all substrates in a surface microstructure‐independent manner. At 7 days, cell numbers were reduced on PT, and DNA synthesis was reduced by 50% on all surfaces. At 7 days, anti‐α5 antibodies caused a partial reduction in alkaline phosphatase enzyme activity on all surfaces, but this effect was independent of surface microstructure. These results indicate that surface micron‐scale topography modulates α5β1 integrin binding and FAK activation. Signaling via α5‐dependent mechanisms is required for DNA synthesis and regulation of alkaline phosphatase, but this effect is independent of surface microstructure. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res, 2007
Electrochemical characterization of albumin protein on Ti‐6AL‐4V alloy immersed in a simulated plasma solution
Authors: Padilla Norma , Bronson Arturo
Published: November 2006

The effect of oxygen and albumin on the electrochemical behavior of a Ti‐6Al‐4V alloy immersed in a simulated inorganic plasma (SIP) solution was studied with a rotating‐cylindrical electrode configuration to focus on the surface/electrolyte reactions. Potentiokinetic scans and electrochemical impedance spectroscopy have been used to characterize the interface by determining the passive current density and capacitance. For the polarization scans, an albumin addition of 37.7 mg/cm3 to the SIP solution (oxygenated and unoxygenated) decreased the passive current density, indicating a lowering of the corrosive rate. The surface capacitance for the Ti‐6Al‐4V alloy immersed in a SIP solution averaged 13 μF/cm2, which transformed after albumin addition (37.7 mg/cm3) from a potential independent behavior to the capacitance ranging from 23 to 6 μF/cm2 with increasing potentials from −800 to 1500 mVSCE, respectively, indicative of albumin adsorption. Within the same potential range and albumin addition to oxygenated solutions, the capacitances expanded slightly with a similar decreasing trend from 31 to 6 μF/cm2, although the capacitance depicts an interaction between the hydrated passive film and the adsorbed albumin from −550 to 500 mVSCE in which the capacitance plateaued at 15 μF/cm2. The hydrated porous oxide film results from the porous rutile layer reacting with H2O2 formed as an intermediary component of oxygen reduction at the Ti‐6Al‐4V surface. The passive film‐albumin interaction would affect the processing of titanium alloys in their surface preparation for biocompatibility, as well as determining the reactivity of titanium alloys to proteins. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res, 2007
Silica–ceramic suppresses the expression of proinflammatory cytokines induced by lipopolysaccharide in macrophages
Authors: Hwang Meeyul , Park Kwan‐Kyu , Chang Young‐Chae , Choo Young‐Ae , Jeon Jae‐Pil , Shin Im‐Hee , Lee Tae‐Sung
Published: November 2006

Bioactive materials have previously been used to coat implants. In a new development for bioactive materials, a silica–ceramic mixture was found to alleviate pain (Lee, Poster presented at the Ninth World Congress of Gynecological Endocrinology, Hongkong, 2001. Poster session (p47)). Here, we hypothesized that silica–ceramic can reduce the expression and activity of cyclooxygenase 2 (COX2) or cytokines associated with inflammation. The production of COX2 and proinflammatory cytokines was investigated by reverse transcriptase (RT)‐PCR and ELISA assay in macrophages stimulated by lipopolysaccharide (LPS). Silica–ceramic had no effect of COX2 expression and prostaglandin production in macrophages. However, silica–ceramic suppressed the synthesis of cytokines involved in inflammation, in particular, the expression of IL‐1β and IL‐6 was reduced at the transcriptional and translational levels. The involvement of NF‐κB in the suppression of cytokines by silica–ceramic was examined by luciferase reporter assay. The NF‐κB activity stimulated by LPS was inhibited by 20–60% with silica–ceramic compared with treatment with LPS alone. We suggest that inhibition of NF‐κB activity by silica–ceramic might cause the attenuation of proinflammatory cytokine expression in macrophages. In conclusion, silica–ceramic could be an alternative approach to regulate the inflammation process. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res, 2007
Suppression effect of seminal vesicle autoantigen on platelet‐activating factor‐induced mouse sperm capacitation
Authors: Huang Yen Hua , Chen Yee Hsiung , Lin Chun Mao , Ciou Yi Yun , Kuo Shin Peih , Chen Chien Tsu , Shih Chwen Ming , Chang E.E.
Published: November 2006

Mammalian sperm gain the ability to fertilize an egg successfully by the capacitation process. An unregulated capacitation process causes sperm to undergo a spontaneous acrosome reaction (AR) and resulting in loss of their fertilization activity. Thus, functional sperm activation is tightly regulated by a capacitation and suppression (decapacitation) mechanism. Factors, such as platelet‐activating factor (PAF) present in both sperm and the female genital tract, are able to stimulate sperm capacitation. Seminal plasma is thought to have the ability to suppress sperm capacitation; however, the regulatory mechanisms of seminal plasma protein on sperm capacitation are not well understood. Recently, we demonstrated that seminal vesicle autoantigen (SVA), a major seminal vesicle secretory protein, is able to suppress mouse sperm capacitation. To further study the suppression spectra of SVA on sperm capacitation, we investigated the effect of SVA on PAF‐induced mouse sperm capacitation‐related signals. Here, we demonstrate that SVA decreases the [Ca2+]i to suppress the PAF's effects on [Ca2+]i, the cAMP level, protein tyrosine phosphorylation, and capacitation. The inhibition of PAF‐induced protein tyrosine phosphorylation and capacitation by SVA can be reversed by cAMP agonists. Characterization of the interactions of SVA with PAF by TLC overlay and tryptophan fluorescence spectrum analyses indicates that SVA is capable of binding PAF with an apparent dissociation constant Kd > 50 µM. Together with these results, we demonstrate that SVA deceases [Ca2+]i and cross‐talks with PAF‐induced intracellular signals to regulate mouse sperm capacitation. J. Cell. Biochem. 100: 941–951, 2007. © 2006 Wiley‐Liss, Inc.
Curcumin prevents lipopolysaccharide‐induced atrogin‐1/MAFbx upregulation and muscle mass loss
Authors: Jin Bingwen , Li Yi‐Ping
Published: November 2006

Because elevated ubiquitin ligase atrogin‐1/MAFbx and MuRF1 mediate skeletal muscle wasting associated with various catabolic conditions, the signaling pathways involved in the upregulation of these genes under pathological conditions are considered therapeutic targets. AKT and NF‐κB have been previously shown to regulate the expression of atrogin‐1/MAFbx or MuRF1, respectively. In addition, we recently found that p38 MAPK mediates TNF‐α upregulation of atrogin‐1/MAFbx expression, suggesting that multiple signaling pathways mediate muscle wasting in inflammatory diseases. To date, however, these advances have not resulted in a practical clinical intervention for disease‐induced muscle wasting. In the present study, we tested the effect of curcumin—a non‐toxic anti‐inflammatory reagent that inhibits p38 and NF‐κB—on lipopolysaccharide (LPS)‐induced muscle wasting in mice. Daily intraperitoneal (i.p.) injection of curcumin (10–60 µg/kg) for 4 days inhibited, in a dose‐dependent manner, the LPS‐stimulated (1 mg/kg, i.p.) increase of atrogin‐1/MAFbx expression in gastrocnemius and extensor digitorum longus (EDL) muscles, resulting in the attenuation of muscle protein loss. It should also be noted that curcumin administration did not alter the basal expression of atrogin‐1/MAFbx, nor did it affect LPS‐stimulated MuRF1 and polyubiquitin expression. LPS activated p38 and NF‐κB, while inhibiting AKT; whereas, curcumin administration inhibited LPS‐stimulated p38 activation, without altering the effect of LPS on NF‐κB and AKT. These results indicate that curcumin is effective in blocking LPS‐induced loss of muscle mass through the inhibition of p38‐mediated upregulation of atrogin‐1/MAFbx. J. Cell. Biochem. 100: 960–969, 2007. © 2006 Wiley‐Liss, Inc.
Gene structure and seasonal expression of carp fish prolactin short receptor isoforms
Authors: San Martín Rody , Hurtado Williams , Quezada Claudia , Reyes Ariel E. , Vera M. Ines , Krauskopf Manuel
Published: November 2006

The complex adaptive mechanisms that eurythermal fish have evolved in response to the seasonal changes of the environment include the transduction of the physical parameter variations into neuroendocrine signals. Studies in carp (Cyprinus carpio) have indicated that prolactin (PRL) and growth hormone (GH) expression is associated with acclimatization, suggesting that the pituitary gland is a relevant physiological node in this adaptive process. Also, the distinctive pattern of expression that carp prolactin receptor (PRLr) protein depicts upon seasonal acclimatization supports the hypothesis that PRL and its receptor clearly are involved in the new homeostatic stage that the eurythermal fish needs to survive during the cyclical changes of its habitat. Here, we characterize the first prolactin receptor gene in a teleost and show that its expression is not associated with alternative promoters, unlike in humans and rodents. Using the regulatory region to direct the transcription of green fluorescent protein (GFP) in zebrafish embryos, we mapped the appearance of this hormone receptor during fish development. This is the first report identifying a fish prolactin receptor gene expressing transcript isoforms encoding for short forms of the protein (45 kDa). These have been found in osmoregulatory tissues of the carp and are regulated in connection with the seasonal acclimatization of the fish. J. Cell. Biochem. 100: 970–980, 2007. © 2006 Wiley‐Liss, Inc.
Exposure to azide markedly decreases the abundance of mRNAs encoding cholesterol synthetic enzymes and inhibits cholesterol synthesis
Authors: Kasturi Sriram , Bederman Ilya R. , Christopher Bridgette , Previs Stephen F. , Ismail‐Beigi Faramarz
Published: November 2006

This study was performed to identify genes that are regulated in the adaptive response to prolonged inhibition of oxidative phosphorylation. Gene microarray analysis in control Clone 9 cells and Clone 9 cells exposed to 5 mM azide for 24 h was carried out as a condition of “Chemical hypoxia.” Among several hundred mRNAs whose abundances were either increased or decreased, we noted that the abundance of mRNAs encoding enzymes that catalyze the sequential steps of cholesterol synthesis was decreased; this finding was verified by real‐time PCR. Exposure to azide for 24 h markedly inhibited the biosynthesis of cholesterol by ∼90% and decreased the cellular content of cholesterol by 30%, similar results were observed in HepG2 cells. The abundance of sterol regulatory element binding protein (SREBP)‐2 mRNA decreased to 0.37 and 0.25 that of controls after 2 and 24 h exposure, respectively. After 24 h of exposure to azide the precursor and nuclear forms of SREBP‐2 protein decreased by ∼80% and ∼50%, respectively. Stimulation of AMP‐activated protein kinase (AMPK) by AICAR in Clone 9 cells increased the abundance of mRNAs encoding cholesterol biosynthetic enzymes and that of SREBP‐1c, and had no effect on SREBP‐2 mRNA abundance. We conclude that the decrease in the abundance of multiple mRNAs encoding cholesterol biosynthetic enzymes may be mediated by decreased expression of SREBP‐2 mRNA and protein and does not involve stimulation of AMPK. The decrease in SREBP‐2 mRNA and protein abundance in the face of decreased cell cholesterol content raises the possibility of a novel regulatory pathway. J. Cell. Biochem. 100: 1034–1044, 2007. © 2006 Wiley‐Liss, Inc.
Role of ROS and MAPK in TPA‐induced ICAM‐1 expression in the myeloid ML‐1 cell line
Authors: Traore Kassim , Sharma Rajni B. , Burek C. Lynne , Trush Michael A.
Published: November 2006

Intercellular adhesion molecule 1 (ICAM‐1) has been implicated in playing a key role in the mechanism of inflammatory process initiated in response to environmental agents, and during normal hematopoietic cell differentiation. Though induction of ICAM‐1 by 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA) in myeloid cells has been reported, the molecular mechanism by which TPA upregulates ICAM‐1 expression remains unclear. In the present study, we investigated the signaling mechanism associated with TPA‐induced ICAM‐1 expression in ML‐1 cells. Herein, our microarray, flow cytometry, and Western blot analysis indicated that ICAM‐1 was constitutively expressed at a low level in ML‐1 cells, but its expression was further upregulated at both the mRNA and protein levels in response to TPA. ICAM‐1 expression in response to TPA was inhibited by pretreatment with GF109203X [a specific inhibitor of protein kinase C (PKC)], or with PD98059 and U0126 (specific inhibitors of MEK), suggesting the importance of PKC, and Erk1/2 signaling cascades in this response. Interestingly, ICAM‐1 expression in response to TPA‐induced PKC activation was linked to the generation of reactive oxygen species (ROS), as pretreatment with NAC (an ROS scavenger) blocked both ErK1/2 activation and ICAM‐1 expression induced by TPA. In addition, TPA‐induced ICAM‐1 expression was blocked by inhibition of nuclear factor‐κB (NF‐κB) activation following pretreatment with BAY11‐7085 (a specific inhibitor of NF‐κB activation). TPA‐induced NF‐κB activation was shown by increased degradation of IkB (NF‐κB specific inhibitory protein). Together, these observations demonstrated that TPA, a potent activator of PKC, induces ICAM‐1 expression via a ROS‐ and ERK1/2‐dependent signaling mechanism in ML‐1 cells. J. Cell. Biochem. 100: 1010–1021, 2007. © 2006 Wiley‐Liss, Inc.
Ovariectomy‐induced bone loss occurs independently of B cells
Authors: Li Yan , Li Aimin , Yang Xiaoying , Weitzmann M. Neale
Published: November 2006

Estrogen withdrawal is associated with a significant expansion in B cell precursor and mature B cell populations. However, despite significant circumstantial evidence the role of B lineage cells in ovariectomy‐induced bone loss in vivo is unclear. In vitro studies have demonstrated that mature B cells have the potential to both positively and negatively impact osteoclastogenesis by virtue of their capacity to secrete pro‐osteoclastogenic cytokines including receptor activator of NFκB ligand (RANKL), as well as anti‐osteoclastogenic cytokines such as osteoprotegerin (OPG) and transforming growth factor beta (TGFβ). Although several studies have suggested that expansion of the B lineage following ovariectomy may play a key role in the etiology of ovariectomy‐induced bone loss, in vivo studies to directly test this notion have yet to be conducted. In this study, we performed ovariectomy on µMT(−/−) mice which are specifically deficient in mature B cells. Analysis of bone mineral density (BMD) by dual‐energy X‐ray absorptiometry (DXA) and micro‐computed tomography (CT) demonstrate that mature B cell‐deficient mice undergo an identical loss of bone mass relative to wild‐type (WT) control mice. Our data demonstrate that mature B cells are not central mediators of ovariectomy‐induced bone loss in vivo. J. Cell. Biochem. 100: 1370–1375, 2007. © 2006 Wiley‐Liss, Inc.
Tristetraprolin recruits functional mRNA decay complexes to ARE sequences
Authors: Hau Heidi H. , Walsh Richard J. , Ogilvie Rachel L. , Williams Darlisha A. , Reilly Cavan S. , Bohjanen Paul R.
Published: November 2006

AU‐rich elements (AREs) in the 3′ untranslated region (UTR) of numerous mammalian transcripts function as instability elements that promote rapid mRNA degradation. Tristetraprolin (TTP) is an ARE‐binding protein that promotes rapid mRNA decay through mechanisms that are poorly understood. A 31 nucleotide ARE sequences from the TNF‐alpha 3′ UTR promoted TTP‐dependent mRNA decay when it was inserted into the 3′ UTR of a beta‐globin reporter transcript, indicating that this short sequence was sufficient for TTP function. We used a gel shift assay to identify a TTP‐containing complex in cytoplasmic extracts from TTP‐transfected HeLa cells that bound specifically to short ARE sequences. This TTP‐containing complex also contained the 5′–3′ exonuclease Xrn1 and the exosome component PM‐scl75 because it was super‐shifted with anti‐Xrn1 or anti‐PMscl75 antibodies. RNA affinity purification verified that these proteins associated specifically with ARE sequences in a TTP‐dependent manner. Using a competition binding assay, we found that the TTP‐containing complex bound with high affinity to short ARE sequences from GM‐CSF, IL‐3, TNF‐alpha, IL‐2, and c‐fos, but did not bind to a U‐rich sequence from c‐myc, a 22 nucleotide poly U sequence or a mutated GM‐CSF control sequence. High affinity binding by the TTP‐containing complex correlated with TTP‐dependent deadenylation and decay of capped, polyadenylated transcripts in a cell‐free mRNA decay assay, suggesting that the TTP‐containing complex was functional. These data support a model whereby TTP functions to enhance mRNA decay by recruiting components of the cellular mRNA decay machinery to the transcript. J. Cell. Biochem. 100: 1477–1492, 2007. © 2006 Wiley‐Liss, Inc.
Sphingosylphosphorylcholine induces apoptosis of endothelial cells through reactive oxygen species‐mediated activation of ERK
Authors: Jeon Eun Su , Lee Mi Jeong , Sung Sang‐Min , Kim Jae Ho
Published: November 2006

Sphingosylphosphorylcholine (SPC) produces reactive oxygen species (ROS) in MS1 pancreatic islet endothelial cells. In the present study, we explored the physiological significance of the SPC‐induced ROS generation in endothelial cells. SPC induced cell death of MS1 cells at higher than 10 µM concentration through a caspase‐3‐dependent pathway. SPC treatment induced sustained activation of an extracellular signal‐regulated kinase (ERK), in contrast to transient activation of ERK in response to platelet‐derived growth factor (PDGF)‐BB, which stimulated proliferation of MS1 cells. Both the SPC‐induced cell death and ERK activation were abolished by pretreatment of the cells with the MEK inhibitor U0126 or by overexpression of a dominant negative mutant of MEK1 (DN‐MEK1). Pretreatment of the cells with N‐acetylcysteine, an antioxidant, completely prevented the SPC‐induced ROS generation, apoptosis, and ERK activation, whereas the ROS generation was not abrogated by treatment with U0126. Consistent with these results, SPC induced cell death of human umbilical vein endothelial cells (HUVECs) through ROS‐mediated activation of ERK. These results suggest that the SPC‐induced generation of ROS plays a crucial role in the cell death of endothelial cells through ERK‐dependent pathway. J. Cell. Biochem. 100: 1536–1547, 2007. © 2006 Wiley‐Liss, Inc.
Ubiquitin‐dependent proteolysis of trihydrophobin 1 (TH1) by the human papilloma virus E6‐associated protein (E6‐AP)
Authors: Yang Yanzhong , Liu Weicheng , Zou Weiying , Wang Hanzhou , Zong Hongliang , Jiang Jianhai , Wang Yanlin , Gu Jianxin
Published: November 2006

Human Papilloma virus E6‐associated protein (E6‐AP), which is known as an E3 ubiquitin ligase, mediates ubiquitination and subsequent degradation of a series of cellular proteins. In this paper, we identify here trihydrophobin 1 (TH1), an integral subunit of the human negative transcription elongation factor (NELF) complex, as a novel E6‐AP interaction protein and a target of E6‐AP‐mediated degradation. Overexpression of E6‐AP results in degradation of TH1 in a dose‐dependent manner, whereas knock‐down of endogenous E6‐AP elevates the TH1 protein level. TH1 protein turnover is substantially faster, compared to controls, in cells that overexpressed E6‐AP. Wild‐type E6‐AP promotes the ubiquitination of TH1, while a catalytically inactive point mutant of E6‐AP abolishes its ubiquitination. Furthermore, in vitro ubiquitination assay also demonstrates that TH1 can be ubiquitinated by E6‐AP. The degradation is blocked by treatment with proteasome inhibitor MG132. Herein, we provide strong evidence that TH1 is a specific substrate that is targeted for degradation through E6‐AP‐catalyzed polyubiquitination. J. Cell. Biochem. 101: 167–180, 2007. © 2006 Wiley‐Liss, Inc.
The experience of HIV/AIDS among rural women in the Northwestern United States: A qualitative analysis
Authors: Mphande‐Finn Joyce T. , Sommers‐Flanagan John
Published: November 2006

The unique experience of HIV/AIDS among rural women in the United
States was explored using qualitative interviews with 7 women who are
HIV positive. Based on these interviews, eight themes emerged. These
included (a) daily powerful emotions, (b) emotional and physical
abandonment, (c) romantic betrayal, (d) medical treatment issues, (e) loss
and grief, (f) appreciating a good support system, (g) renewed purpose for
living, and (h) personal growth and transformation. Results are discussed
with regard to future research and practice with rural women with
HIV/AIDS, with an emphasis on the importance of social support and
counseling opportunities in facilitating women's progress through the
emotional process associated with HIV/AIDS diagnosis and treatment.
© 2007 Wiley Periodicals, Inc.
Contributors to assessments of risk in intimate partner violence: How victims and professionals differ
Authors: Cattaneo Lauren Bennett
Published: November 2006

This article explores what factors contribute to victims' vs. victim advocates' assessments of risk of repeat intimate partner violence. A sample of 169 court‐involved victims and the advocates who conducted intake with them were asked to rate victims' risk of repeat abuse. Significant contributors to victim assessments were her level of post‐traumatic stress disorder symptoms, the batterer's general violence, whether she had been living with the batterer at the time of the offense, and the level of psychological abuse in the relationship. For advocates, significant contributors were the batterer's drug use, whether the batterer and victim had children in common, and the level of physical and psychological abuse in the relationship. Implications of these differences and directions for future study are discussed. © 2007 Wiley Periodicals, Inc.
The depiction of mental illnesses in children's television programs
Authors: Wahl Otto , Hanrahan Erin , Karl Kelly , Lasher Erin , Swaye Janel
Published: November 2006

Concern has been expressed that negative attitudes toward people with mental illnesses begin to develop early in childhood. This study examines one of the possible sources of learning of such negative attitudes—children's television programs. Two hundred sixty‐nine (269) hours of children's television programming were videotaped, viewed, and rated. Nearly half the programs viewed contained use of mental health terms. Most of these terms were slang and disrespectful (e.g., crazy and nut) and were used to disparage others. Twenty‐ one (21) characters with a mental illness were identified. These characters tended to be depicted as aggressive and threatening, and other characters responded to them with fear, exclusion, and disrespect. Children's television programs seem to provide substantial opportunity for children to develop negative attitudes and behaviors toward those with mental illnesses. © 2007 Wiley Periodicals, Inc.