bluuebunny/crossref_metadata_2025 huggingface dataset:
Adenoviral-mediated expression of Pcsk9 in mice results in a low-density lipoprotein receptor knockout phenotype
Authors: Maxwell Kara N. , Breslow Jan L.
Published: April 2004

Proprotein convertase subtilisin kexin 9 (Pcsk9) is a subtilisin serine protease with a putative role in cholesterol metabolism. Pcsk9 expression is down-regulated by dietary cholesterol, and mutations in Pcsk9 have been associated with a form of autosomal dominant hypercholesterolemia. To study the function of Pcsk9 in mice, an adenovirus constitutively expressing murine Pcsk9 (Pcsk9-Ad) was used. Pcsk9 overexpression in wild-type mice caused a 2-fold increase in plasma total cholesterol and a 5-fold increase in non-high-density lipoprotein (HDL) cholesterol, with no increase in HDL cholesterol, as compared with mice infected with a control adenovirus. Fast protein liquid chromatography analysis showed that the increase in non-HDL cholesterol was due to an increase in low-density lipoprotein (LDL) cholesterol. This effect appeared to depend on the LDL receptor (LDLR) because LDLR knockout mice infected with Pcsk9-Ad had no change in plasma cholesterol levels as compared with knockout mice infected with a control adenovirus. Furthermore, whereas overexpression of Pcsk9 had no effect on LDLR mRNA levels, there was a near absence of LDLR protein in animals overexpressing Pcsk9. These results were confirmed
in vitro
by the demonstration that transfection of Pcsk9 in McA-RH7777 cells caused a reduction in LDLR protein and LDL binding. In summary, these results indicate that overexpression of Pcsk9 interferes with LDLR-mediated LDL cholesterol uptake. Because Pcsk9 and LDLR are coordinately regulated by cholesterol, Pcsk9 may be involved in a novel mechanism to modulate LDLR function by an alternative pathway than classic cholesterol inhibition of sterol regulatory element binding protein-mediated transcription.
Adaptor Protein Complex 1 Mediates the Transport of Lysosomal Proteins from a Golgi-like Organelle to Peripheral Vacuoles in the Primitive EukaryoteGiardia lamblia
Authors: Touz María C. , Kulakova Liudmila , Nash Theodore E.
Published: April 2004

Giardia lamblia is an early branching protist that possesses peripheral vacuoles (PVs) with characteristics of lysosome-like organelles, located underneath the plasma membrane. In more evolved cells, lysosomal protein trafficking is achieved by cargo recognition involving adaptor protein (AP) complexes that recognize specific amino acid sequences (tyrosine and/or dileucine motifs) within the cytoplasmic tail of membrane proteins. Previously, we reported that Giardia has a tyrosine-based sorting system, which mediates the targeting of a membrane-associated cysteine protease (encystation-specific cysteine protease, ESCP) to the PVs. Here, we show that Giardia AP1 mediates the transport of ESCP and the soluble acid phosphatase (AcPh) to the PVs. By using the yeast two-hybrid assay we found that the ESCP tyrosine-based motif interacts specifically with the medium subunit of AP1 (Giμa). Hemagglutinin-tagged Giμa colocalizes with ESCP and AcPh and coimmunoprecipitates with clathrin, suggesting that protein trafficking toward the PVs is clathrin-adaptin dependent. Targeted disruption of Giμa results in mislocalization of ESCP and AcPh but not of variant-specific surface proteins. Our results suggest that, unlike mammalian cells, only AP1 is involved in anterograde protein trafficking to the PVs in Giardia. Moreover, even though Giardia trophozoites lack a morphologically discernible Golgi apparatus, the presence of a clathrin-adaptor system suggests that this parasite possess a primitive secretory organelle capable of sorting proteins similar to that of more evolved cells.
Bid-Cardiolipin Interaction at Mitochondrial Contact Site Contributes to Mitochondrial Cristae Reorganization and CytochromecRelease
Authors: Kim Tae-Hyoung , Zhao Yongge , Ding Wen-Xing , Shin Jin Na , He Xi , Seo Young-Woo , Chen Jun , Rabinowich Hannah , Amoscato Andrew A. , Yin Xiao-Ming
Published: April 2004

Release of cytochrome c from the mitochondrial intermembrane space is critical to apoptosis induced by a variety of death stimuli. Bid is a BH3-only prodeath Bcl-2 family protein that can potently activate this efflux. In the current study, we investigated the mitochondrial localization of Bid and its interactions with mitochondrial phospholipids, focusing on their relationships with Bid-induced cytochrome c release. We found that Bid binding to the mitochondria required only three of its eight helical structures (α4-α6), but not the BH3 domain, and the binding could not be inhibited by the antideath molecule Bcl-xL. Membrane fractionations indicated that tBid bound to mitochondrial outer membranes at both contact and noncontact sites. Bid could interact with specific cardiolipin species on intact mitochondria as identified by mass spectrometry. Like the binding to the mitochondria, this interaction could not be blocked by the mutation in the BH3 domain or by Bcl-xL.However, a cardiolipin-specific dye, 10-N-nonyl acridine orange, could preferentially suppress Bid binding to the mitochondrial contact site and inhibit Bid-induced mitochondrial cristae reorganization and cytochrome c release. These findings thus suggest that interactions of Bid with mitochondrial cardiolipin at the contact site can contribute significantly to its functions.
A multi‐dimensional view of entrepreneurship
Authors: Yamada Jin‐ichiro
Published: April 2004

This paper attempts to synthesise the theoretical research on entrepreneurship and social capital undertaken in previous studies, and presents a multi‐dimensional view of entrepreneurship. In examining overviews of past single perspective entrepreneurship research, this study shows that the primary role of entrepreneurs in organisation emergence is to acquire knowledge and create social capital properly. This process is necessarily accompanied by creation of knowledge communities to establish the domain consensus of new organisations among various stakeholders paying particular attention to the fragility and dysfunctional side of entrepreneurship and social capital. The paper concludes with a hypothesis and suggestions towards a future research agenda. It is hoped that, as a result of this theoretical development, this paper will help to focus greater attention on the concept of entrepreneurship in studies of management development.
Consulting as story‐making
Authors: Johansson Anders W.
Published: April 2004

Story‐telling has been identified as a metaphor for management consulting which focuses on the consultant as “impression manager”. The convincing power of strong stories told by consultants is emphasised. In this article management consulting is conceptualised through the metaphor of story‐making, identifying co‐operation and reflection as characteristic features. A specific consultancy process is interpreted as a co‐operative effort between the consultant and the client in which plot lines and characters are constructed. Plots can be opened or closed, and characters can be settled or negotiated. Thus story‐making as a reflective act moves between enhancing reflection and preparing for action.
Strategic learning in Germany's largest companies
Authors: Hilse Heiko , Nicolai Alexander T.
Published: April 2004

Over the past few years, corporate universities, as they are called, have become increasingly common. Independent of this, strategy process theory is also finding a place in international management research. In this paper, it will be shown that strategy process theory demonstrates an affinity with the concept of the corporate university. A survey of the 1,000 largest German companies for the “Federal Ministry of Education and Research” is examining for the first time how widespread corporate universities are in Germany and how they are organized. On the basis of this empirical data it will be shown to what extent the concept of the corporate university in practice is in line with the theoretical findings of strategy process theory.
Assessing intercultural training designs
Authors: Graf Andrea
Published: April 2004

Training designs are critical to the success of intercultural training programmes. A common typology for classifying intercultural training designs distinguishes among the following dimensions: experiential discovery versus didactic expository and culture‐specific versus culture‐general training. The purpose of this paper is to assess different intercultural training designs referring to this typology. First, a concept of intercultural competence is developed. Based on this concept experiential training designs are favoured as they allow to train the cognitive, affective and behavioural component of intercultural competencies. In the second part of the paper an empirical study in the USA and Germany is conducted in order to assess whether the degree of intercultural competencies differs between the two nations. As no considerable differences can be identified the benefit of culture‐general training is discussed. Significant skills to be focused on in intercultural training are described and specific measures for pre‐assessment devices in intercultural training are presented.
The role of brand name in customization decisions: a search vs experience perspective
Authors: Jiang Pingjun
Published: April 2004

Investigates the role of brand name in consumers' decision making during a customization process, and develops a conceptual understanding of the factors influencing the role of brand name from a “search vs experience” perspective. Addresses the strategic relationship of brand with perceived product/service/information preference match and the impact of preference match on consumer “willingness to pay for customization”. Brand name was found to be an important decision variable for customization in terms of getting a better preference match. Brand name still holds an important role on consumer communications, as was expected. Therefore, it is reasonable to believe that customization would not “commoditize” brands, but rather increase the effect of brand names in purchase decision making. High‐knowledge consumers reported stronger brand name effect in terms of its importance and usefulness in their decision making. The brand name effect varies across product categories, and the effect is stronger in the customization of search products than that in experience products. Thus, brand names have greater impact on choices in a search product where less total quality information on components is available for facilitating consumer choices. Discusses managerial implications of the study's findings.
The personal brand in futures
Authors: Hines Andy
Published: April 2004

Considers branding more in line with bringing forth self‐knowledge and self‐expression than the creation of a cultivated or false self. It's not about selling “new and improved” futures tools. Rather, it's about discovering the inner and authentic you and proclaiming it to your clients. The personal brand is about recognizing who you are and what you do best, and communicating that to clients.
A first record of Isistius plutodus in the north‐eastern Atlantic
Authors: Zidowitz H. , Fock H. O. , Pusch C. , Von Westernhagen H.
Published: April 2004

One specimen of the largetooth cookiecutter shark Isistius plutodus was caught in the north‐eastern Atlantic at 43°58′ N; 28°32′ W. This is the first record of this rare species in the north‐eastern Atlantic and the northernmost point of its known distribution.
Identification and mapping of a rice blast resistance gene Pi‐g(t) in the cultivar Guangchangzhan
Authors: Zhou J. H. , Wang J. L. , Xu J. C. , Lei C. L. , Ling Z. Z.
Published: April 2004

A single dominant blast resistance gene was identified in Chinese indica rice (Oryza sativa) cv. Guangchangzhan (GCZ), which shows complete resistance to Japanese isolate Ken53‐33 of Magnaporthe grisea. Genetic analysis of the backcross (BC1) and second‐generation (F2) populations from a cross between susceptible cv. Lijiangxintuanheigu (LTH) and GCZ indicated that the resistance was conferred by one dominant gene. This gene was mapped on the long arm of chromosome 2 and flanked by RM166 and RM208 at distances of 4·00 ± 4·90 and 6·30 ± 4·89 cM (centiMorgans), respectively. It was designated tentatively as Pi‐g(t).
Water sorption and the plasticization effect in wafers
Authors: Martínez‐Navarrete Nuria , Moraga Gemma , Talens Pau , Chiralt Amparo
Published: April 2004

Wafers are low moisture food products whose crunchiness or crispness is considered a primary textural attribute, highly affected by the product's physical state (glassy or rubbery). The water activity–water content–glass transition relationships for commercial wafers are reported, using Gordon and Taylor's equation to model the water plasticization effect and also Brunnaver‐Emmett‐Teller (BET) and Guggenheim‐Anderson‐DeBoer (GAB) sorption models. BET monolayer moisture content was 6.2% and the moisture limit to fit this model was about 11.5%. Critical water activity and critical water content for the glass transition of the product at 20 °C were 0.591 and 0.118 (mass fraction) respectively. Abrupt changes in the mechanical product properties, evaluated from a three‐point bend test, could be observed at these limits. Moisture levels between 6 and 11% give rise to a glassy state matrix in the product and so to acceptable product crispness. Below this range, the glassy matrix seems to turn fragile and, above this range, the product becomes rubbery.
Tectono‐metamorphic evolution of the Okcheon Metamorphic Belt, South Korea: Tectonic implications in East Asia
Authors: Oh Chang Whan , Kim Sung Won , Ryu In‐Chang , Okada Toshinori , Hyodo Hironobu , Itaya Tetsumaru
Published: April 2004

Abstract  The tectonic history of the Okcheon Metamorphic Belt (OMB) is a key to understanding the tectonic relationship between South Korea, China and Japan. The petrochemistry of 150 psammitic rocks in the OMB indicates that the depositional environment progressively deepened towards the northwest. These data, combined with the distribution pattern of oxide minerals and the abundance of carbonaceous material, support a half‐graben basin model for the OMB. Biotite and muscovite K–Ar dates from metasediments in the central OMB range from 102 to 277 Ma. K–Ar ages of 142–194 Ma are widespread throughout the area, whereas the older ages of 216–277 Ma are restricted to the metasediments of the middle part of the central OMB. The younger (Cretaceous) ages are only found in metasediments that are situated near the Cretaceous granite intrusions. The 216–277 Ma dates from weakly deformed areas represent cooling ages of M1 intermediate pressure/temperature (P/T) metamorphism. The relationship between age distribution and deformation pattern indicates that the Jurassic muscovite and biotite dates can be interpreted as complete resetting ages, caused by thermal and deformational activities associated with Jurassic granite plutonism. Well‐defined 40Ar/39Ar plateau ages of 155–169 Ma for micas from both metasediments and granitic rocks can be correlated with the main Jurassic K–Ar mica ages (149–194 Ma). U–Pb zircon dates for biotite granite from the southwest OMB are 167–169 Ma. On the basis of the predominantly Jurassic igneous and metamorphic ages and the uniformity of d002 values for carbonaceous materials in the study area, it is suggested that the OMB has undergone amphibolite facies M2 metamorphism after M1 metamorphism. This low P/T M2 regional thermal metamorphism may have been caused by the regional intrusion of Jurassic granites. The OMB may have undergone tectono‐metamorphic evolution as follows: (i) the OMB was initiated as an intraplate rift in the Neoproterozoic during break‐up of Rodinia, and may represent the extension of Huanan aulacogen within the South China block; (ii) sedimentation continued from the Neoproterozoic to the Ordovician, perhaps with several unconformities; (iii) M1 intermediate P/T metamorphism occurred during the Late Paleozoic due to compression caused by collision between the North and South China blocks in an area peripheral to the collision zone; and (iv) during the Early to Middle Jurassic, north‐westward subduction of the Farallon‐Izanagi Plate under the Asian Plate resulted in widespread intrusion of granites, which triggered M2 low P/T regional thermal metamorphism in the OMB. This event also formed the dextral Honam shear zone at the boundary between the OMB and Precambrian Yeongnam massif.
Catching Up and Convergence: Long‐run Growth in East Asia
Authors: Hsiao Frank S.T. , Hsiao Mei‐Chu W.
Published: April 2004

The paper attempts to combine the traditional learning model with the recent theory of economic growth using Maddison's long‐run real GDP per capita data of the three fastest growing countries in East Asia: Korea, Taiwan, and Japan. The authors first explain games of catching‐up among nations, and then explain the learning coefficients of Taiwan and Korea with Japan and the United States through periods before and after World War II. The model of leaning leads to the logistic model of economic growth of convergence between two countries. Using time‐series data, the coefficients of a logistic model are estimated to confirm that the real GDP per capita of Taiwan and Korea are converging to that of Japan and the United States, respectively. Similarly, Japan's GDP per capita converges to that of the United States. The time required for finite convergence for these countries is also estimated.
International Trade and Gender Wage Discrimination: Evidence from East Asia
Authors: Berik Günseli , Rodgers Yana van der Meulen , Zveglich Joseph E.
Published: April 2004

The paper explores how competition from international trade affects gender wage discrimination in two open economies. According to neoclassical theory, if discrimination is costly, then increased industry competitiveness from international trade lessens the incentive for employers to discriminate against women. This effect should be stronger in concentrated sectors, where employers can use excess profits to cover the costs of discrimination. Alternatively, increased international trade may reduce women's bargaining power to achieve wage gains. Results for Taiwan and Korea indicate that, in contrast to neoclassical theory, competition from foreign trade in concentrated industries is positively associated with wage discrimination against women.
On the Economics of Refugee Flows
Authors: Stark Oded
Published: April 2004

Although the phenomenon of refugee flows is not devoid of economic connotations, it has so far been investigated primarily by political scientists and sociologists. The analytical tools of economic inquiry have not yet been applied to this subject, although it stands to reason that such a study will contribute to our understanding of why refugee flows occur and will guide the policy response. This note illustrates how economic analysis can be brought to bear on three key aspects of refugee flows: fear, poverty, and group movement.
Coping in marital dyads: A contextual perspective on the role of gender and health
Authors: Badr Hoda
Published: April 2004

In order to create a single measure that simultaneously measures emotion‐, problem‐, and relationship‐focused coping, an exploratory factor analysis was performed onCoyne and Smith's (1994)Relationship‐Focused Coping Scale (RFCS) andCarver's (1997)Brief COPE, using data collected from 182 married couples. In 90 couples both spouses were healthy; in 92 couples, one spouse had a chronic illness. Results yielded an abridged 23‐item, 6‐factor measure. Using the new measure, a series of repeated measures analyses of variance were conducted to determine whether coping styles vary by gender and health. Results showed husbands and wives differ in their use of active engagement, approach coping, and protective buffering depending on whether they are healthy or ill. Guided byKenny's (1990)Actor‐Partner Interaction Model (APIM), results of hierarchical linear modeling analyses using SAS Proc Mixed showed couples who are more congruent in their use of active engagement, and more complementary in their use of protective buffering and avoidance coping report greater marital adjustment. Although husbands and wives may employ different coping strategies, these results highlight the importance of examining the ways in which spouses copetogetherin the face of a shared stressor.

The course of true love never did run smooth.–

—William Shakespeare
Predicting Participation in Premarital Prevention Programs: The Health Belief Model and Social Norms
Authors: Sullivan Kieran T. , Pasch Lauri A. , Cornelius Tara , Cirigliano Ellen
Published: April 2004

The development of effective programs to prevent marital dysfunction has been a recent focus for marital researchers, but the effective dissemination of these programs to engaged couples has received relatively little attention. The purpose of this study is to determine which factors predict couples’ participation in premarital counseling. Predictive factors were derived from the health prevention literature, with a particular focus on the health belief model (HBM). Couples’ beliefs and attitudes about premarital counseling were assessed at least six months before their wedding, and participation was assessed after their wedding. Results indicate that the strongest predictors of couples’ participation were couples’ perceptions of barriers to counseling and whether they had counseling recommended to them. These variables predicted participation even after controlling for important demographic variables. Recommendations for recruiting engaged couples for premarital counseling are made based on the findings.
Interest in and Barriers to Participation in Multiple Family Groups Among Head and Neck Cancer Survivors and Their Primary Family Caregivers
Authors: Ostroff Jamie , Ross Stephanie , Steinglass Peter , Ronis‐tobin Victor , Singh Bhuvanesh
Published: April 2004

This study examined interest in and barriers to participation in a multiple family group intervention (MFG) for adult cancer survivors and their family caregivers. The intervention was developed to assist families in coping with the persistent challenges of cancer diagnosis, treatment, and rehabilitation. Eighty eligible families having a member diagnosed and treated for cancers of the head and neck region completed a baseline quality of life survey consisting of standardized psychosocial measures, and then all patients and their families were invited to participate in a day‐long multiple family group program. However, despite extensive recruitment efforts and accommodations to address anticipated barriers for nonparticipation, only 15 of the 80 (19%) eligible families agreed to attend the MFG workshop. Post‐MFG, participating families reported high levels of program satisfaction and usefulness. These findings are discussed in the context of increasing the use of family‐focused interventions in cancer care settings.
Sacred Practices in Highly Religious Families: Christian, Jewish, Mormon, and Muslim Perspectives1
Authors: Marks Loren
Published: April 2004

Quantitative research examining linkages between family relationships and religious experience has increased substantially in recent years. However, related qualitative research, including research that examines the processes and meanings behind recurring religion‐family correlations, remains scant. To address this paucity, a racially diverse sample (N=24) of married, highly religious Christian, Jewish, Mormon, and Muslim parents of school‐aged children were interviewed regarding the importance of religious family interactions, rituals, and practices in their families. Mothers and fathers discussed several religious practices that were meaningful to them and explained why these practices were meaningful. Parents also identified costs and challenges associated with these practices. Interview data are presented in connection with three themes: (1) “practicing [and parenting] what you preach,” (2) religious practices, family connection, and family communion, and (3) costs of family religious practices. The importance of family clinicians and researchers attending to the influence of religious practice in the lives of highly religious individuals and families is discussed.
Coeval Large-Scale Magmatism in the Kalahari and Laurentian Cratons During Rodinia Assembly
Authors: Hanson Richard E. , Crowley James L. , Bowring Samuel A. , Ramezani Jahandar , Gose Wulf A. , Dalziel Ian W. D. , Pancake James A. , Seidel Emily K. , Blenkinsop Thomas G. , Mukwakwami Joshua
Published: April 2004

We show that intraplate magmatism occurred 1106 to 1112 million years ago over an area of two million square kilometers within the Kalahari craton of southern Africa, during the same magnetic polarity chron as voluminous magmatism within the cratonic core of North America. These contemporaneous magmatic events occurred while the Rodinia supercontinent was being assembled and are inferred to be parts of a single large igneous province emplaced across the two cratons. Widespread intraplate magmatism during Rodinia assembly shows that mantle upwellings required to generate such provinces may occur independently of the supercontinent cycle.
Immunogenicity of Constrained Monoclonal Antibody A32-Human Immunodeficiency Virus (HIV) Env gp120 Complexes Compared to That of Recombinant HIV Type 1 gp120 Envelope Glycoproteins
Authors: Liao Hua-Xin , Alam S. Munir , Mascola John R. , Robinson James , Ma Benjiang , Montefiori David C. , Rhein Maria , Sutherland Laura L. , Scearce Richard , Haynes Barton F.
Published: April 2004

One strategy for the generation of broadly reactive neutralizing antibodies (NA) against human immunodeficiency virus type 1 (HIV-1) primary isolates is to use immunogens that have constrained HIV-1 envelope gp120 conformations reflective of triggered envelope on the surface of virions. A major change in gp120 following binding to CD4 is the enhanced exposure of the CCR5 binding site. One inducer of CCR5 binding site epitopes on gp120 is the human anti-gp120 monoclonal antibody, A32. We have made cross-linked A32-rgp120
89.6
and A32-rgp120
BaL
complexes and have compared their immunogenicities to those of uncomplexed recombinant gp120
BaL
(rgp120
BaL
) and rgp120
89.6
. A32-rgp120
89.6
and A32-rgp120
BaL
complexes had stable induced CCR5 binding site expression compared to that of uncomplexed rgp120s. However, the A32-rgp120 complexes had similar capacities in guinea pigs for induction of NA against HIV-1 primary isolates versus that of rgp120 alone. A32-rgp120
89.6
induced antibodies that neutralized 6 out of 11 HIV-1 isolates, while rgp120
89.6
alone induced antibodies that neutralized 4 out of 11 HIV-1 isolates. A32-rgp120
BaL
complexes induced antibodies that neutralized 4 out of 14 HIV-1 isolates while, surprisingly, non-cross-linked rgp120
BaL
induced antibodies that neutralized 9 out of 14 (64%) HIV-1 isolates. Thus, stable enhanced expression of the coreceptor binding site on constrained gp120 is not sufficient for inducing broadly neutralizing anti-HIV-1 NA. Moreover, the ability of HIV-1 rgp120
BaL
to induce antibodies that neutralized ∼60% of subtype B HIV-1 isolates warrants consideration of using HIV-1 BaL as a starting point for immunogen design for subtype B HIV-1 experimental immunogens.
Analysis of Adenovirus Sequestration in the Liver, Transduction of Hepatic Cells, and Innate Toxicity after Injection of Fiber-Modified Vectors
Authors: Shayakhmetov Dmitry M. , Li Zong-Yi , Ni Shaoheng , Lieber André
Published: April 2004

After intravenous administration, adenovirus (Ad) vectors are predominantly sequestered by the liver. Delineating the mechanisms for Ad accumulation in the liver is crucial for a better understanding of Ad clearance and Ad-associated innate toxicity. To help address these issues, in this study, we used Ad vectors with different fiber shaft lengths and either coxsackievirus-Ad receptor (CAR)-interacting Ad serotype 9 (Ad9) or non-CAR-interacting Ad35 fiber knob domains. We analyzed the kinetics of Ad vector accumulation in the liver, uptake into hepatocytes and Kupffer cells, and induction of cytokine expression and release in response to systemic vector application. Immediately after intravenous injection, all Ad vectors accumulated equally efficiently in the liver; however, only genomes of long-shafted Ads were maintained in the liver tissue over time. We found that Kupffer cell uptake of long-shafted Ads was mediated by the fiber knob domain and was CAR independent. The short-shafted Ads were unable to efficiently interact with hepatocellular receptors and were not taken up by Kupffer cells. Moreover, our studies indicated that Kupffer cells were not the major reservoir for the observed accumulation of Ads (used in this study) in the liver within the first 30 min after virus infusion. The lower level of liver cell transduction by short-shafted Ads correlated with a significantly reduced inflammatory anti-Ad response as well as liver damage induced by the systemic administration of these vectors. This study contributes to a better understanding of the biology of systemically applied Ad and will help in designing safer vectors that can efficiently transduce target tissues.
Receptor-Dependent Coronavirus Infection of Dendritic Cells
Authors: Turner Brian C. , Hemmila Erin M. , Beauchemin Nicole , Holmes Kathryn V.
Published: April 2004

In several mammalian species, including humans, coronavirus infection can modulate the host immune response. We show a potential role of dendritic cells (DC) in murine coronavirus-induced immune modulation and pathogenesis by demonstrating that the JAW SII DC line and primary DC from BALB/c mice and p/p mice with reduced expression of the murine coronavirus receptor, murine CEACAM1a, are susceptible to murine coronavirus infection by a receptor-dependent pathway.
Focal nodular hyperplasia with concomitant hepatocellular carcinoma: a case report and clonal analysis
Authors: Zhang S-H , Cong W-M , Wu M-C
Published: April 2004

This report describes a hepatocellular carcinoma (HCC) with concomitant focal nodular hyperplasia (FNH) in a 56 year old Chinese man. There were two well circumscribed tumours measuring 3 × 2.5 × 2 cm and 2 × 1.5 × 1.5 cm. The larger mass was grey and soft with a small area of bleeding and necrosis and an intact capsule. The smaller mass was yellow and had no capsule. Clonal analysis was carried out to clarify the relation between the HCC and the adjacent FNH. The clonal analysis was based on the methylation pattern of the polymorphic X chromosome linked androgen receptor gene (HUMARA). In FNH, after HpaII digestion, the allelic bands showed two well defined peaks. The intensity of the two peaks in the DNA from cirrhotic tissue did not differ significantly, consistent with a random pattern of X chromosome inactivation. However, in HCC, after HpaII digestion, the allelic bands differed significantly in intensity. Therefore, there was a typical polyclonal pattern of inactivation in FNH but the HCC was interpreted as being monoclonal.
NGF gene expression and secretion in white adipose tissue: regulation in 3T3-L1 adipocytes by hormones and inflammatory cytokines
Authors: Peeraully Muhammad R. , Jenkins John R. , Trayhurn Paul
Published: April 2004

The sympathetic nervous system plays a central role in lipolysis and the production of leptin in white adipose tissue (WAT). In this study, we have examined whether nerve growth factor (NGF), a target-derived neurotropin that is a key signal in the development and survival of sympathetic neurons, is expressed and secreted by white adipocytes. NGF mRNA was detected by RT-PCR in the major WAT depots of mice (epididymal, perirenal, omental, mesenteric, subcutaneous) and in human fat (subcutaneous, omental). In mouse WAT, NGF expression was observed in mature adipocytes and in stromal vascular cells. NGF expression was also evident in 3T3-L1 cells before and after differentiation into adipocytes. NGF protein, measured by ELISA, was secreted from 3T3-L1 cells, release being higher before differentiation. Addition of the sympathetic agonists norepinephrine, isoprenaline, or BRL-37344 (β3-agonist) led to falls in NGF gene expression and secretion by 3T3-L1 adipocytes, as did IL-6 and the PPARγ agonist rosiglitazone. A substantial decrease in NGF expression and secretion occurred with dexamethasone. In contrast, LPS increased NGF mRNA levels and NGF secretion. A major increase in NGF mRNA level (9-fold) and NGF secretion (≤40-fold) in 3T3-L1 adipocytes occurred with TNF-α. RT-PCR showed that the genes encoding the p75 and trkA NGF receptors were expressed in mouse WAT. These results demonstrate that white adipocytes secrete NGF (an adipokine), NGF synthesis being influenced by several factors with TNF-α having a major stimulatory effect. We suggest that NGF is a target-derived neurotropin and an inflammatory response protein in white adipocytes.
Transcriptional regulation of the lactase-phlorizin hydrolase promoter by PDX-1
Authors: Wang Zhi , Fang Rixun , Olds Lynne C. , Sibley Eric
Published: April 2004

Lactase-phlorizin hydrolase gene expression is spatially restricted along the anterior-posterior gut axis. Lactase gene transcription is maximal in the distal duodenum and jejunum in adult mammals and is barely detectable in the proximal duodenum. By contrast, pancreatic duodenal homeobox-1 (PDX-1) protein is expressed maximally in the proximal duodenum. This study aimed to determine the role of PDX-1 in regulating lactase gene promoter activity in intestinal epithelial cells. Caco-2 cells were cotransfected with lactase promoter-reporter constructs in the presence of a PDX-1 expression vector and assayed for luciferase activity. PDX-1 cotransfection results in repression of lactase promoter activity. Sequence analysis of the lactase promoter revealed a putative PDX-1 DNA binding site in the proximal 100-bp lactase gene promoter. EMSAs demonstrated that PDX-1 can interact with the lactase promoter binding site but not with a site in which the core PDX-1 binding sequence TAAT is mutated. Site-directed mutagenesis of the PDX-1 core binding site in the lactase promoter-reporter construct suggests that PDX-1 can function independently of DNA binding to its consensus binding site. Stable overexpression of PDX-1 results in repression of the endogenous human lactase gene in differentiated Caco-2 cells. Given the contrasting spatial expression pattern, PDX-1 may function to specify the anterior boundary of lactase expression in the small intestine and is thus a candidate regulator of anterior spatial restriction in the gut.
Creatine kinase-deficient hearts exhibit increased susceptibility to ischemia-reperfusion injury and impaired calcium homeostasis
Authors: Spindler Matthias , Meyer Klaus , Strömer Hinrik , Leupold Andrea , Boehm Ernest , Wagner Helga , Neubauer Stefan
Published: April 2004

The creatine kinase (CK) system is involved in the rapid transport of high-energy phosphates from the mitochondria to the sites of maximal energy requirements such as myofibrils and sarcolemmal ion pumps. Hearts of mice with a combined knockout of cytosolic M-CK and mitochondrial CK (M/Mito-CK−/−) show unchanged basal left ventricular (LV) performance but reduced myocardial high-energy phosphate concentrations. Moreover, skeletal muscle from M/Mito-CK−/−mice demonstrates altered Ca2+homeostasis. Our hypothesis was that in CK-deficient hearts, a cardiac phenotype can be unmasked during acute stress conditions and that susceptibility to ischemia-reperfusion injury is increased because of altered Ca2+homeostasis. We simultaneously studied LV performance and myocardial Ca2+metabolism in isolated, perfused hearts of M/Mito-CK−/−( n = 6) and wild-type (WT, n = 8) mice during baseline, 20 min of no-flow ischemia, and recovery. Whereas LV performance was not different during baseline conditions, LV contracture during ischemia developed significantly earlier (408 ± 72 vs. 678 ± 54 s) and to a greater extent (50 ± 2 vs. 36 ± 3 mmHg) in M/Mito-CK−/−mice. During reperfusion, recovery of diastolic function was impaired (LV end-diastolic pressure: 22 ± 3 vs. 10 ± 2 mmHg), whereas recovery of systolic performance was delayed, in M/Mito-CK−/−mice. In parallel, Ca2+transients were similar during baseline conditions; however, M/Mito-CK−/−mice showed a greater increase in diastolic Ca2+concentration ([Ca2+]) during ischemia (237 ± 54% vs. 167 ± 25% of basal [Ca2+]) compared with WT mice. In conclusion, CK-deficient hearts show an increased susceptibility of LV performance and Ca2+homeostasis to ischemic injury, associated with a blunted postischemic recovery. This demonstrates a key function of an intact CK system for maintenance of Ca2+homeostasis and LV mechanics under metabolic stress conditions.
In situ dynamically monitoring the proteolytic function of the ubiquitin-proteasome system in cultured cardiac myocytes
Authors: Dong Xin , Liu Jinbao , Zheng Hanqiao , Glasford Joseph W. , Huang Wei , Chen Quan Hai , Harden Niels R. , Li Faqian , Gerdes A. Martin , Wang Xuejun
Published: April 2004

The ubiquitin-proteasome system (UPS) is responsible for turnover of most cellular proteins in eukaryotes. Protein degradation by the UPS serves quality control and regulatory functions. Proteasome inhibition showed great promise in effectively treating cancer and restenosis. UPS dysfunction in cardiac hypertrophy and failure has recently been suspected but remains to be investigated. A system capable of monitoring dynamic changes in proteolytic function of the UPS in cardiac myocytes in situ would no doubt benefit significantly efforts to decipher the pathogenic significance of UPS dysfunction in the heart and to evaluate the effect of proteasome inhibition on cardiac myocytes. We successfully established such a system in cultured cardiac myocytes by delivering and expressing a modified green fluorescence protein (GFPu) gene using recombinant adenoviruses. GFPu contains a ubiquitination signal sequence fused to the COOH terminus. Fluorescence microscopy and Western blots revealed that protein abundance of modified green fluorescent protein (GFPu), but not wild-type green fluorescent protein, in cultured cardiac myocytes was incrementally increased when function of the proteasomes was inhibited in various degrees by specific inhibitors. The increase in GFPu protein levels and fluorescence intensity is paralleled by a decrease in the in vitro peptidase activity of the proteasomes. Our results demonstrate that GFPu can be used as a surrogate marker to monitor dynamic changes in proteolytic function of the UPS in cardiac myocytes in situ. Application of this novel system reveals that moderate levels of H2O2, a reactive oxygen species generator, impair proteolytic function of the UPS in cultured cardiac myocytes.
Angiotensin II inhibits NaCl absorption in the rat medullary thick ascending limb
Authors: Lerolle Nicolas , Bourgeois Soline , Leviel Françoise , Lebrun Gaëtan , Paillard Michel , Houillier Pascal
Published: April 2004

NaCl reabsorption in the medullary thick ascending limb of Henle (MTALH) contributes to NaCl balance and is also responsible for the creation of medullary interstitial hypertonicity. Despite the presence of angiotensin II subtype 1 (AT1) receptors in both the luminal and the basolateral plasma membranes of MTALH cells, no information is available on the effect of angiotensin II on NaCl reabsorption in MTALH and, furthermore, on angiotensin II-dependent medullary interstitial osmolality. MTALHs from male Sprague-Dawley rats were isolated and microperfused in vitro; transepithelial net chloride absorption ( JCl) as well as transepithelial voltage ( Vte) were measured. Luminal or peritubular 10−11and 10−10M angiotensin II had no effect on JClor Vte. However, 10−8M luminal or peritubular angiotensin II reversibly decreased both JCland Vte. The effect of both luminal and peritubular angiotensin II was prevented by the presence of losartan (10−6M). By contrast, PD-23319, an AT2-receptor antagonist, did not alter the inhibitory effect of 10−8M angiotensin II. Finally, no additive effect of luminal and peritubular angiotensin II was observed. We conclude that both luminal and peritubular angiotensin II inhibit NaCl absorption in the MTALH via AT1receptors. Because of intrarenal angiotensin II synthesis, angiotensin II concentration in medullary tubular and interstitial fluids may be similar in vivo to the concentration that displays an inhibitory effect on NaCl reabsorption under the present experimental conditions.
Lipoxygenase-dependent superoxide release in skeletal muscle
Authors: Zuo Li , Christofi Fievos L. , Wright Valerie P. , Bao Shengying , Clanton Thomas L.
Published: April 2004

Superoxide anion radical (O2•−) is released from skeletal muscle at rest and is particularly elevated during conditions of heat stress (42°C). Previous studies have shown that in isolated rat diaphragm O2•−release is not dependent on mitochondrial electron transport, reduced NADP oxidase activity, or the integrity of membrane anion channels. This study hypothesized that O2•−release, as measured by cytochrome c reduction, is linked to metabolism of arachidonic acid. Phospholipase A2inhibition with manoalide significantly decreased O2•−release. In downstream pathways, neither the blockage of cyclooxygenase with indomethacin nor the inhibition of cytochrome P-450-dependent monooxygenase with SKF-525A decreased O2•−release. However, lipoxygenase (LOX) inhibition with general LOX blockers 5,8,11,14-eicosatetraynoic acid and cinnamyl-3,4-dihydroxy-α-cyanocinnamate greatly attenuated the signal. Furthermore, the specific 5-LOX inhibitor diethylcarbamazine also significantly decreased O2•−release. Immunohistochemistry localized 5- and 12-LOX to the cytosol and sarcolemma of muscle cells. Confocal studies, using the O2•−-sensitive fluorescent indicator hydroethidine, demonstrated that LOX inhibition had no significant influence on intracellular O2•−formation. When compared with the cytochrome c results, this indicates that intra- and extracellular O2•−must arise from different sources. These data show for the first time that arachidonic acid metabolism through LOX activity, is a major source of extracellular O2•−release in skeletal muscle.
What random variable generates a bounded potential?
Authors: Kartashov N. , Mishura Yu.
Published: April 2004

It is known that if a predictable nondecreasing process generates
a bounded potential, then its final value satisfies the Garsia
inequality. We prove the converse, that is, a random
variable satisfying the Garsia inequality generates a bounded
potential. We also propose some useful relations between the
Garsia inequality and the Cramer conditions, and different ways
how to construct a potential.
Essential 𝒰cκ‐type maps andBirkhoff‐Kellogg theorems
Authors: Agarwal R. P. , O′Regan Donal
Published: April 2004

We present a new continuation theorem for
‐type maps. The analysis is elementary
and relies on properties of retractions and fixed point theory
for self‐maps. Also we present some Birkhoff‐Kellogg type
theorems on invariant directions.
Murine Model of Neointimal Formation and Stenosis in Vein Grafts
Authors: Cooley Brian C.
Published: April 2004

Objective—

Previous studies have suggested that neointimal formation, a central cause of vein graft stenosis, has several potential cell sources. It was hypothesized that neointimal cells arise primarily from the cells of the vein graft.

Methods and Results—

This study investigated vein graft neointimal cell origins using a model of vein-to-artery cross-transplantation between transgenic Rosa26 mice (constitutive expression of bacterial β-galactosidase marker gene) and wild-type mice. Vein-originating cells survived and make a major contribution to neointimal formation within the vein graft, mostly adjacent to the lumen/endothelium, suggesting an intimate association with endothelial cells. Cross-transplantation of veins from thrombomodulin promoter-driven β-galactosidase reporter transgenic mice to wild-type arteries demonstrated survival of vein graft endothelial cells. Neointimal thickening was greater at the proximal and, to a lesser extent, distal ends, in comparison to the middle of the graft. By contrast, arterial grafts had almost no neointimal formation throughout the graft. The relative neointimal wall thickness is much greater in this model compared with other murine and larger-species vein graft models, even showing near-occlusive stenosis of the perianastomotic region.

Conclusions—

Vein graft neointimal cells arise predominantly from vein-derived cells, suggesting clinical relevance of stenosis-inhibiting therapies directed at the vein graft.
Malonyl Coenzyme A Decarboxylase Inhibition Protects the Ischemic Heart by Inhibiting Fatty Acid Oxidation and Stimulating Glucose Oxidation
Authors: Dyck Jason R.B. , Cheng Jie-Fei , Stanley William C. , Barr Rick , Chandler Margaret P. , Brown Steven , Wallace David , Arrhenius Thomas , Harmon Charles , Yang Guang , Nadzan Alex M. , Lopaschuk Gary D.
Published: April 2004

Abnormally high rates of fatty acid oxidation and low rates of glucose oxidation are important contributors to the severity of ischemic heart disease. Malonyl coenzyme A (CoA) regulates fatty acid oxidation by inhibiting mitochondrial uptake of fatty acids. Malonyl CoA decarboxylase (MCD) is involved in the decarboxylation of malonyl CoA to acetyl CoA. Therefore, inhibition of MCD may decrease fatty acid oxidation and protect the ischemic heart, secondary to increasing malonyl CoA levels. Ex vivo working rat hearts aerobically perfused in the presence of newly developed MCD inhibitors showed an increase in malonyl CoA levels, which was accompanied by both a significant decrease in fatty acid oxidation rates and an increase in glucose oxidation rates compared with controls. Using a model of demand-induced ischemia in pigs, MCD inhibition significantly increased glucose oxidation rates and reduced lactate production compared with vehicle-treated hearts, which was accompanied by a significant increase in cardiac work compared with controls. In a more severe rat heart global ischemia/reperfusion model, glucose oxidation was significantly increased and cardiac function was significantly improved during reperfusion in hearts treated with the MCD inhibitor compared with controls. Together, our data show that MCD inhibitors, which increase myocardial malonyl CoA levels, decrease fatty acid oxidation and accelerate glucose oxidation in both ex vivo rat hearts and in vivo pig hearts. This switch in energy substrate preference improves cardiac function during and after ischemia, suggesting that pharmacological inhibition of MCD may be a novel approach to treating ischemic heart disease.