bluuebunny/crossref_metadata_2025 huggingface dataset:
Organising the Academy: New Organisational Forms and the Future of the University
Authors: Jacob Merle , Hellström Tomas
Published: October 2003

Decreasing public spending, new accountability measures and calls for broader ranges of services to society have combined to encourage the reorganisation of European universities. Universities are asked to restructure themselves along lines suggested by corporate management. This paper examines this proposal by looking at four management models in the light of new and traditional universities. The paper discusses the principle of ‘heterarchical organising’, which has emerged spontaneously within the university, and argues that this appears to be a more appropriate model for university organisation.
The Survival of the Democratic Intellect: Academic Values in Scotland and England
Authors: Paterson Lindsay
Published: October 2003

Using the results of a sample survey of academics in higher education institutions in Scotland and England, the paper assesses attitudes to the civic role of higher education. It places these in the context of debates about core academic values, about the public accountability of higher education institutions, and about the devolution of political power in the United Kingdom. It finds that there is widespread attachment to a civic role for higher education, alongside strong attachment to traditional academic values. These values are not significantly influenced by individual academics’ gender, age or social class of origin, and differences by academic discipline are not as strong as might be expected. Academics in Scotland tend to hold a somewhat more civic view than academics in England. This national difference seems to be a product of distinctive national systems, since academics of English origin in Scotland share in the majority Scottish views.
Budgetary Control in a Decentralized System: Meeting the Criteria for Fiscal Stability in the European Union
Authors: Wildavsky Aaron , Jones L.R.
Published: October 2003

This article provides a set of measures that are recommended to the leadership of the European Union to enhance budgetary balance and control within a highly decentralized system, one that requires the coordination of budgetary policy in more than a dozen nations. These measures are intended to be relevant to European budgetary coordination efforts but also appear to apply to U.S. federal initiatives to reduce the annual budget deficit and total debt. The individual measures recommended include holding increases in expenditures at or below the rate of GDP growth, establishing envelopes for major accounts, placing all entitlement on a pay‐as‐you‐go basis, means testing of entitlements and elimination of automatic entitlement indexing, considering the option of marginal reductions in some entitlement programs, and voting on budgets as single, omnibus packages. The proposal to establish executive line‐item budget authority also is evaluated‐and rejected.
Performance Measurement in a Direct Democratic Environment: Local Government Reforms in Switzerland
Authors: Schedler Kuno
Published: October 2003

Current international development in public management is characterized by a rare unanimity among countries. Trends in public administration, business administration, applied economics, and public law indicate a stronger emphasis on performance and a weakening of input‐oriented analysis and steering. At the foundation of this development is performance measurement (PM). However, the political and administrative circumstance of its implementation differ widely. The following article deals with the implications of Swiss direct democracy for new approaches to public management and, specifically, performance measurement.
Local Government Finance in Hungary: Okun Revisited
Authors: Straussman Jeffrey D. , Fabian Katalin
Published: October 2003

Fundamental change in the economic systems of the nations‐of central Europe is now taking place. The economic transition has broad social and cultural consequences, among these the alteration of local government service provision and financial management. In this article, the authors describe some of the fiscal characteristics of local governments in Hungary with an analysis of the dilemmas of transition. Efforts to create and enhance local government financial management capability are detailed. This article portrays the difficulty of developing a fully operational market economy within a democratic context in the post‐Communist era while, at the same time, maintaining social protection for citizens through the continued provision of a wide array of social benefit programs. The essential dilemma for local government social program managers and budget officials is that service demand far surpasses the financial resources available to continue services at the levels to which people grew accustomed under the Communist regime.
The Evolution of Local Government Budgeting in Poland: From Accounting to Policy in a Leap and a Bound
Authors: Thurmaier Kurt
Published: October 2003

The evolution of U.S. public budgeting has been described by Schick (1966) as a progression from a control orientation to a management and then a planning orientation. Lee (1992) has tracked the evolution of central budgeting staff largely from a group of accountants to a more varied set of backgrounds, dominated by public administration and general liberal arts educations. As Poland decentralizes fiscal and administrative responsibilities and powers to gminas (local self‐governments), the gmina councils are beginning to view the budget as more than a financial accounting tool proscribed and prescribed by the central government. They are beginning to see the potential for using the budget as a policy and management tool. The evolutionary transformation of budgeting, which took fifty years in the United States, may take only five years in Poland. The evidence for this metamorphosis is based on interviews with several gmina city budgeters, with special attention devoted to the city of Lublin.
Mexican Economic and Fiscal Policies to Sustain Political Stability
Authors: Cothran Dan A.
Published: October 2003

During the 1980s, Mexico was in a state of crisis. Its economic prosperity collapsed in 1982, and its political system almost self‐destructed in 1988. Yet by 1994, the economy was once again growing and the party that had ruled the country since 1929 appeared poised to win the national election of August 1994, despite the traumatic assassination of the ruling party candidate for president. How did the Mexican regime do it? This article examines the role of economic policies in restoring the fortunes of the Mexican regime.
Why Is Central Regulation of Local Spending Decisions So Pervasive? Evidence From a Case Study
Authors: Carlsen Fredrik
Published: October 2003

While the “classical” theory of fiscal federalism argues that local authorities should have substantial discretion to set the local budgetary mix, surveys show that central regulation of local spending decisions is pervasive. Norway provides an ideal setting for studying spending regulations. An attempt to decentralize fiscal choice to the local levels, the block grant reform, has been reversed in recent years. This article presents a case study of the interaction between a county and the national government from 1986, when the reform was implemented, to 1992. The study concludes that the government introduced new regulations to prevent local authorities from distorting spending decisions in order to elicit higher grants.
Reporting on the Control Structures of Local Government Under the Single Audit Act of 1984
Authors: Jakubowski Stephen
Published: October 2003

One of the major objectives of the Single Audit Act of 1984 (SAA) is to improve the financial management of local governments (Public Law 98–502, 98 Stat.2327). The purpose of this study is to examine how local governments have responded to the SAA and to provide some evidence concerning the extent to which the SAA is accomplishing its stated objectives. This study examines what auditors have reported with respect to the control structures of local governments and considers the extent to which control structures changed over the first four reporting years under the SAA. Differences in the frequency of reported weaknesses are examined across the four years and between types of government‐counties and cities‐and types of auditors‐ large and small public accounting firms and state auditors. The study also considers the extent to which reported weaknesses were corrected. The research findings reveal that auditors reported considerably more control weaknesses in counties than in cities, and the frequency of control weaknesses reported declined significantly over the first four reporting years under the SAA for city governments but remained relatively constant for county governments. In addition, there were significant differences in the frequency of control weaknesses reported related to the type of auditor.
Government Risk‐Financing Pools: An Assessment of Current Practices
Authors: Young Peter C. , Reed B.J.
Published: October 2003

Since 1970, state and local governments have experienced two “insurance crises;” the first occurred in the mid‐ to late‐1970s and the second in the mid‐80s. The result has been a twenty‐year period of time in which state and local governments have been able to afford insurance only intermittently‐if insurance has been available at all. In response to this problem, local governments, government associations, and state governments created alternative risk‐finartcing mechanisms to provide coverage for themselves. These mechanisms, commonly referred to as self‐insurance pools, enable local governments within a state to pool together risks and resources to finance the costs of fortuitous losses. In 1988, the first comprehensive examination of pooling practices was undertaken through a nationwide survey. This article reports the findings from the first follow‐up study since the 1988 survey and further extends knowledge of pooling and pooling practices.
After the Earthquake: The President's Budget for FY 1996
Authors: Caiden Naomi
Published: October 2003

What changes has the political earthquake of November 1994 wreaked on the federal budget and, in particular, on the president's budget? Are the policy dynamics of federal budgets so entrenched that they transcend politics and are impervious to shocks? Or do the political changes presage radical policy changes, reversing decades of cumulative commitments? Are budgetary institutions and processes flexible enough to accommodate enhanced conflicts while maintaining their integrity and purpose? This article discusses the fate of the 1995 budget and the initial problems faced in the president's budget for 1996.
Is The National Budget Controllable?
Authors: True James L.
Published: October 2003

Previous scholarly analyses of national spending have shown a penchant for incre‐mentalism in interpreting changes. They have also focused almost exclusively on annual national outlays or annual expenditures. This article argues that, in many cases, budget authority figures provide a better basis for analyzing the national budget. The author gathered annual budget authority (BA) figures from fiscal years 1969 to 1993 and used this BA data for an empirical test of budget controllability. If the national budget has or has not been controlled in the recent past, then it should help us to judge whether or not it is controllable now. Five potential major budget interventions from fiscal year 1980 to 1993 were tested using an interrupted time‐series and two regression models. Different intervention results are anticipated depending upon whether one has a policy perspective that is incrementalist, international systemic, or domestic political. The analysis provides more support for the domestic political perspective than for incrementalist or international systemic views. Budget authority was found to be superior to outlays for linking budget results to policy decisions. The national budget responded appropriately to policy decisions in the recent past. In this sense, the budget was controlled and we should view it as controllable.
Performance‐Based Government Models: Building A Track Record
Authors: Broom Cheryle A.
Published: October 2003

Governments across the country have embarked on performance‐based government efforts. Typical goals of these efforts are to clarify the mission and priority objectives of government with an emphasis on the expected results, to develop mechanisms for monitoring and reporting the achievement of those objectives, and to use this information to make decisions about government activities, including making government more accountable Although many governments have reported success in establishing their performance‐based programs, we found ourselves wondering whether any of these programs are showing signs that they can be sustained over time. Are performance‐based reports or budgets being used by managers and policy makers? Are the performance programs maintained after implementation? If so, what are the factors critical to success?

To answer these questions, we reviewed performance‐based government efforts in progress throughout the country with a focus on the initiatives of five states.1 This discussion begins by describing the efforts in those five states: the stated need, the approach, and the results to date. We then discuss the elements of success and the lessons learned from these performance‐based government pioneers.
Defense Business Operations Fund (DBOF): Problems and Promise
Authors: Jordan Leland G.
Published: October 2003

In 1991, the United States Department of Defense established the Defense Business Operations Fund (DBOF). The DBOF includes support activities with a business volume well in excess of $80 billion annually, and it provides activities and services that are essential to the operation of the military forces. The necessity to reduce the defense budget and to find innovative ways of producing more defense than would be possible under prior operating and management arrangements caused the Department of Defense (DoD) to look to alternative economic and business models.

The changes made are dramatic. The implementation has not gone as smoothly as some observers expected, and Congress and former Secretary of Defense Aspin each considered reversing the innovation.

This article addresses the problems and the promised improvements in the context of the economic and management concepts undergirding the DBOF, concepts that are dramatically different than those previously applied in the DoD
Earmarking as a Strategy Against Budgetary Constraints: San Francisco's Children's Amendment
Authors: Stowers Genie N.L.
Published: October 2003

The passage and implementation of the San Francisco Children's Amendment, or Proposition J, in 1991 represents an important expansion of the increasingly common strategy of budgetary earmarking by human service organizations. The core of the initiative was a city charter amendment that earmarked a portion of existing property taxes and established a baseline budget for children's services. Of these provisions, the baseline budget has proven most important in preserving children's services and preventing any substitution effects in a time of increasing fiscal stress for the city/ county of San Francisco.

This case also illustrates the role of a strong political appeal in passing technical budgetary initiatives over the objections of elected officials and others. While there was concern about the reduced flexibility created by earmarking a city's budget during a period of retrenchment, this concern was overlooked because of the overwhelming political support for the children's cause.

However, conflict between groups was created by the measure as groups fought to be included in the provisions of the amendment and over whether each group had obtained its fair share of funding.
State Tax Cuts of 1995: Is Something New Afoot?
Authors: Gold Steven D.
Published: October 2003

Perhaps the hottest fiscal topic in state capitals in 1995 was tax cuts. When legislatures convened in January, it appeared possible that states were on the verge of an unprecedented wave of tax cuts. The Republican landslide of November 1994 swept into office many new governors and legislators who had campaigned on reducing taxes. Governor Christine Todd Whitman of New Jersey was considered a role model by many of them. Perhaps state tax policy was about to take a major new direction.

According to some reports, that is precisely what happened. For example, Stephen Moore, Director of Fiscal Policy Studies at the Cato Institute, wrote the following introduction to his overview of state tax policies:1

1995 is shaping up to be the biggest tax‐cutting year in more than 10 years…. [T]he supply side philosophy that low tax rates and expenditure controls are the key fiscal tools to state economic competitiveness now appears to be the new governing philosophy in state capitals from Albany to Sacramento. The Pete Wilson, Lowell Weicker, Jim Florio soak‐the‐rich philosophy of the early 1990s has been nearly universally repudiated across the states. Meanwhile, “Whitmanomics,” named after tax‐cutting Christine Todd Whitman of New Jersey, is spreading rapidly to other states.

In fact, there was a wave of state tax cuts in 1995 with about half of the states enacting reductions of some kind. But it appears premature to draw the strong conclusion that this marked a radical change in state fiscal policy. State legislatures trimmed or rejected most of the proposals for large tax cuts. Most of the 1995 tax cuts were small or modest in size Rather than a new departure, they are consistent with past policies at similar points in the business cycle.

It is also possible, however, that something new really is afoot. Conservative anti‐tax forces are gaining strength in the states, just as they are at the federal level. If it were not for prospective federal aid cuts, state tax policy really might become more conservative. But the shift of federal responsibilities to states will represent a formidable obstacle to large state tax cuts in the coming years.

This article places the tax cuts of 1995 in perspective. It begins with a detailed discussion of ambiguities'in defining and measuring tax reductions and increases. This discussion can be applied to any time period and will remain relevant after concern
The Impact of the Tax Revolt Era State Fiscal Caps: A Research Update
Authors: King‐Meadows Tyson , Lowery David
Published: October 2003

Fiscal caps, the most common form of fiscal limit adopted during the tax revolt era, are again on the agendas of state government. In this article, we evaluate the claims made by cap supporters and opponents by examining the impacts of caps adopted during the tax revolt. Updating Lowery and Cox's (1990) analysis of the impact of state fiscal caps through 1991 using a comparative state, interrupted time‐series design, we find some evidence—albeit very weak—that fiscal caps may have modestly reduced the size of government and no evidence that they have been evaded through budget end‐runs.
Confessions of a Dinosaur
Authors: Kliman Albert J.
Published: October 2003

The old time budget officer has been all but wiped out—the species having been deprived of its natural habitat by the great smoke cloud that began to spread after the great cataclysm of 1974 when the Budget Act began to change all the rules. This, of course, is an overstatement. But it is clear—at least from the perspective of this old dinosaur—that the job of the budget officer has changed over the years, from what many perceived of as a master of financial magic to just another serf (or supervisor) in the bureaucratic feudal system.

This is not a polemic against the Budget Act or the changes that have taken place. The only thing constant about the budget process is change. Rather, it is a discussion of how these changes have affected budget officers over the decades that I have been involved in the budget business.
Ventilatory responses and carotid body function in adult rats perinatally exposed to hyperoxia
Authors: Prieto‐Lloret J. , Caceres A. I. , Obeso A. , Rocher A. , Rigual R. , Agapito M. T. , Bustamante R. , Castañeda J. , Perez‐Garcia M. T. , Lopez‐Lopez J. R. , Gonzalez C.
Published: October 2003

Hypoxia increases the release of neurotransmitters from chemoreceptor cells of the carotid body (CB) and the activity in the carotid sinus nerve (CSN) sensory fibers, elevating ventilatory drive. According to previous reports, perinatal hyperoxia causes CSN hypotrophy and varied diminishment of CB function and the hypoxic ventilatory response. The present study aimed to characterize the presumptive hyperoxic damage. Hyperoxic rats were born and reared for 28 days in 55%–60% O2; subsequent growth (to 3.5–4.5 months) was in a normal atmosphere. Hyperoxic and control rats (born and reared in a normal atmosphere) responded with a similar increase in ventilatory frequency to hypoxia and hypercapnia. In comparison with the controls, hyperoxic CBs showed (1) half the size, but comparable percentage area positive to tyrosine hydroxylase (chemoreceptor cells) in histological sections; (2) a twofold increase in dopamine (DA) concentration, but a 50% reduction in DA synthesis rate; (3) a 75% reduction in hypoxia‐evoked DA release, but normal high [K+]0‐evoked release; (4) a 75% reduction in the number of hypoxia‐sensitive CSN fibers (although responding units displayed a nearly normal hypoxic response); and (5) a smaller percentage of chemoreceptor cells that increased [Ca2+]1 in hypoxia, although responses were within the normal range. We conclude that perinatal hyperoxia causes atrophy of the CB–CSN complex, resulting in a smaller number of chemoreceptor cells and fibers. Additionally, hyperoxia damages O2‐sensing, but not exocytotic, machinery in most surviving chemoreceptor cells. Although hyperoxic CBs contain substantially smaller numbers of chemoreceptor cells/sensory fibers responsive to hypoxia they appear sufficient to evoke normal increases in ventilatory frequency.
Phosphoarginine regulation of the squid nerve Na+/Ca2+ exchanger: metabolic pathway and exchanger–ligand interactions different from those seen with ATP
Authors: DiPolo Reinaldo , Berberián Graciela , Beaugé Luis
Published: October 2003

In squid nerves the Na+–Ca2+ exchanger is up‐regulated by ATP and phosphoarginine (PA). ATP regulation involves drastic alterations in the Na+i, H+i and Ca2+i interactions with the large intracellular cytoplasmic loop of the exchanger protein. In this work we explored the mechanisms associated with PA regulation in intracellular dialysed squid axons and squid optic nerve membrane vesicles. Dialysed axons were used to measure the four modes of exchange fluxes (Na+o–Ca2+i or forward exchange, Ca2+o–Na+i or reverse exchange, Ca2+o–Ca2+i exchange and Na+o–Na+i exchange) under controlled intra‐ and extracellular conditions. Inside‐out membrane vesicles allowed measurement of the Na+‐gradient‐dependent 45Ca2+ uptake (forward mode) as influenced by ligands and digestion with chymotrypsin from the intracellular side. The results show that, unlike ATP, PA regulation does not affect the H+i, Na+i and Ca2+i interactions with the intracellular ‘regulatory’ loop, but increases the affinity of the intracellular transport sites, preferentially for Ca2+i (about 20‐fold) over Na+i (50%); i.e. PA favours the forward mode over the other exchange modes. Intracellular chymotrypsin digestion removed ATP regulation while leaving modulation by PA unmodified. Western blot analysis suggested that chymotrypsin disrupts the large intracellular loop. Together these results indicate that ATP and PA regulations are associated with different structures inside and outside the exchanger protein. Based on these observations we expanded our previous model for metabolic regulation of the Na+–Ca2+ exchanger by adding to the original ‘ATP region’ a new zone, the ‘PA region’, related to the intracellular transport sites for Na+i and Ca2+i. This new model is able to explain most previous and present results.
Transference of recombinant VE‐cadherin cytoplasmic domain alters endothelial junctional integrity and porcine microvascular permeability
Authors: Guo Mingzhang , Wu Mack H. , Granger Harris J. , Yuan Sarah Y.
Published: October 2003

VE‐cadherin constitutes endothelial adherens junctions through a homophilic binding of its extracellular domain and by the anchoring of its intracellular domain to actin cytoskeleton via catenins. The aim of this study was to determine the functional importance of VE‐cadherin–cytoskeleton association in the maintenance of endothelial junctional integrity. A recombinant VE‐cadherin cytoplasmic domain (rVE‐cad CPD) was expressed in E. coli and purified through Ni‐NTA spin columns. Immunoprecipitation assays showed that rVE‐cad CPD was able to bind β‐catenin in vitro and to compete with endogenous VE‐cadherin for binding of β‐catenin in human umbilical vein endothelial cells. A significant increase in the transendothelial flux of albumin was observed in the endothelial cell monolayers transfected with rVE‐cad CPD. Importantly, transfection of rVE‐cad CPD into intact isolated coronary venules markedly elevated the albumin permeability of the venular endothelium. In addition, immunofluorescence microscopic analysis revealed a conformational change of VE‐cadherin from a uniform, continuous distribution along the cell membrane under control conditions to a diffuse, stitch‐like pattern after rVE‐cad CPD transfection. The effects were likely due to an attenuated anchorage of endogenous VE‐cadherin to the cytoskeleton, as evidenced by a decreased partitioning of VE‐cadherin in the detergent‐insoluble cytoskeletal pool. The results suggest that the intracellular association of VE‐cadherin with β‐catenin‐linked cytoskeleton is essential to the maintenance of endothelial junctional integrity and microvascular permeability.
Glycine rectifies vascular dysfunction induced by dietary protein imbalance during pregnancy
Authors: Brawley L. , Torrens C. , Anthony F. W. , Itoh S. , Wheeler T. , Jackson A. A. , Clough G. F. , Poston L. , Hanson M. A.
Published: October 2003

Protein restriction in rat pregnancy programmes the development of elevated systolic blood pressure and vascular dysfunction in the offspring. A recent study has shown that hypertension is reversed by maternal glycine supplementation. Whether this protective effect is exerted directly on the embryo and fetus, or indirectly via effects on the mother, is unknown although we have previously shown abnormalities in the maternal vasculature. We tested the hypothesis that dietary glycine repletion would reverse endothelial dysfunction in protein‐restricted pregnant rat dams using wire myography. Impaired acetylcholine‐ (P < 0.01) and isoprenaline‐induced (P < 0.05) vasodilatation in isolated mesenteric arteries (MA) from protein‐restricted pregnant dams was accompanied by reduced vascular nitric oxide (NO) release (P < 0.05). Dietary glycine supplementation reversed vascular dysfunction in MA (P < 0.05) and improved NO release thus potentially protecting the maternal circulation. The impaired NO release in the MA of low protein diet dams was not accompanied by reduced eNOS mRNA expression, suggesting that eNOS activity was altered. Protein restriction did not alter the vascular function of a conduit artery, the thoracic aorta. These results provide evidence that adequate provision of glycine, a conditionally essential amino acid in pregnancy, may play a role in the vascular adaptations to pregnancy, protecting the fetus from abnormal programming of the cardiovascular system.
Functional expression of TASK‐1/TASK‐3 heteromers in cerebellar granule cells
Authors: Kang Dawon , Han Jaehee , Talley Edmund M. , Bayliss Douglas A. , Kim Donghee
Published: October 2003

TASK‐1 and TASK‐3 are functional members of the tandem‐pore K+ (K2P) channel family, and mRNAs for both channels are expressed together in many brain regions. Although TASK‐1 and TASK‐3 subunits are able to form heteromers when their complementary RNAs are injected into oocytes, whether functional heteromers are present in the native tissue is not known. Using cultured cerebellar granule (CG) neurones that express mRNAs of both TASK‐1 and TASK‐3, we studied the presence of heteromers by comparing the sensitivities of cloned and native K+ channels to extracellular pH (pHo) and ruthenium red. The single‐channel conductance of TASK‐1, TASK‐3 and a tandem construct (TASK‐1/TASK‐3) expressed in COS‐7 cells were 14.2 ± 0.4, 37.8 ± 0.7 and 38.1 ± 0.7 pS (–60 mV), respectively. TASK‐3 and TASK‐1/TASK‐3 (and TASK‐3/TASK‐1) displayed nearly identical single‐channel kinetics. TASK‐3 and TASK‐1/TASK‐3 expressed in COS‐7 cells were inhibited by 26 ± 4 and 36 ± 2 %, respectively, when pHo was changed from 8.3 to 7.3. In outside‐out patches from CG neurones, the K+ channel with single channel properties similar to those of TASK‐3 was inhibited by 31 ± 7 % by the same reduction in pHo. TASK‐3 and TASK‐1/TASK‐3 expressed in COS‐7 cells were inhibited by 78 ± 7 and 3 ± 4 %, respectively, when 5 μm ruthenium red was applied to outside‐out patches. In outside‐out patches from CG neurones containing a 38 pS channel, two types of responses to ruthenium red were observed. Ruthenium red inhibited the channel activity by 77 ± 5 % in 42 % of patches (range: 72–82 %) and by 5 ± 4 % (range: 0–9 %) in 58 % of patches. When patches contained more than three 38 pS channels, the average response to ruthenium red was 47 ± 6 % inhibition (n= 5). These electrophysiological studies show that native 38 pS K+ channels of the TASK family in cultured CG neurones consist of both homomeric TASK‐3 and heteromeric TASK‐1/TASK‐3.
Cortisol enhances structural maturation of the hypoplastic fetal lung in sheep
Authors: Boland Rochelle , Joyce Belinda J. , Wallace Megan J. , Stanton Heather , Fosang Amanda J. , Pierce Richard A. , Harding Richard , Hooper Stuart B.
Published: October 2003

Although exogenous corticosteroids advance structural maturation of the fetal lung, they can adversely affect fetal lung and body growth. Our aim was to determine whether cortisol, at physiological doses, can enhance structural maturation of the hypoplastic fetal lung without affecting fetal lung growth. Fetal sheep were divided into four groups (n= 5 for each) and lung hypoplasia (LH) was induced in two groups. Increasing doses of cortisol (1.5–4.0 mg) were infused into one group of fetuses with LH and one group without LH; the other two groups received saline. LH retarded structural development, reduced tropoelastin mRNA levels, reduced hydroxyproline and elastin contents, and increased active matrix metalloproteinase‐2 (MMP‐2) levels in the fetal lung. Cortisol infusions had no effect on fetal lung growth or body weights. In fetuses with LH, cortisol increased the percentage airspace, reduced the interalveolar wall thickness, increased alveolar number and reduced the increase in active MMP‐2 levels. Thus, relatively low doses of cortisol can enhance structural maturation of the fetal lung without adversely affecting fetal lung growth. However, cortisol did not correct the abnormal deposition of elastin within the alveolar parenchyma associated with LH, indicating that secondary septal crest formation remained abnormal.
Electrophysiological properties of human mesenchymal stem cells
Authors: Heubach Jürgen F. , Graf Eva M. , Leutheuser Judith , Bock Manja , Balana Bartosz , Zahanich Ihor , Christ Torsten , Boxberger Sabine , Wettwer Erich , Ravens Ursula
Published: October 2003

Human mesenchymal stem cells (hMSC) have gained considerable interest due to their potential use for cell replacement therapy and tissue engineering. One strategy is to differentiate these bone marrow stem cells in vitro into cardiomyocytes prior to implantation. In this context ion channels can be important functional markers of cardiac differentiation. At present there is little information about the electrophysiological behaviour of the undifferentiated hMSC. We therefore investigated mRNA expression of 26 ion channel subunits using semiquantitative RT‐PCR and recorded transmembrane ion currents with the whole‐cell voltage clamp technique. Bone marrow hMSC were obtained from healthy donors. The cells revealed a distinct pattern of ion channel mRNA with high expression levels for some channel subunits (e.g. Kv4.2, Kv4.3, MaxiK, HCN2, and α1C of the L‐type calcium channel). Outward currents were recorded in almost all cells. The most abundant outward current rapidly activated at potentials positive to +20 mV. This current was identified as a large‐conductance voltage‐ and Ca2+‐activated K+ current, conducted by MaxiK channels, due to its high sensitivity to tetraethylammonium (IC50= 340 μm) and its inhibition by 100 nm iberiotoxin. A large fraction of cells also demonstrated a more slowly activating current at potentials positive to –30 mV. This current was selectively inhibited by clofilium (IC50= 0.8 μm). Ba2+ inward currents, stimulated by 1 μm BayK 8644 were found in a few cells, indicating the expression of functional L‐type Ca2+ channels. Other inward currents such as sodium currents or inward rectifier currents were absent. We conclude that undifferentiated hMSC express a distinct pattern of ion channel mRNA and functional ion channels that might contribute to physiological cell function.
Regulation of NF-κB-Dependent Lymphocyte Activation and Development by Paracaspase
Authors: Ruefli-Brasse Astrid A. , French Dorothy M. , Dixit Vishva M.
Published: October 2003

Paracaspase (MALT1), a member of an evolutionarily conserved superfamily of caspase-like proteins, has been shown to bind and colocalize with the protein Bcl10 in vitro and, because of this association, has been suggested to be involved in the CARMA1-Bcl10 pathway of antigen-induced nuclear factor κB (NF-κB) activation. We demonstrate that primary T and B lymphocytes from paracaspase-deficient mice are defective in antigen-receptor–induced NF-κB activation, cytokine production, and proliferation. Paracaspase acts downstream of Bcl10 to induce NF-κB activation and is required for the normal development of B cells, indicating that paracaspase provides the missing link between Bcl10 and activation of the IκB kinase complex.
Antibody-Mediated Protection against Cytotoxic T-Cell Escape in Coronavirus-Induced Demyelination
Authors: Dandekar Ajai A. , Jacobsen Gary , Waldschmidt Thomas J. , Perlman Stanley
Published: October 2003

C57BL/6 (B6) mice infected with mouse hepatitis virus (MHV) strain JHM develop a clinically evident, demyelinating encephalomyelitis. Infectious virus can be isolated from the spinal cords of these mice and is invariably mutated in the immunodominant CD8 T-cell epitope recognized in this strain. We showed previously that these persistently infected mice did not mount a measurable serum anti-MHV neutralizing antibody response. Here we show that cytotoxic T-lymphocyte (CTL) escape was not detected in MHV-infected BALB/b mice (H-2bhaplotype), even though the same CD8 T-cell epitopes were recognized as in B6 mice. BALB/b mice had 25-fold more MHV-specific antibody-secreting cells in the central nervous system, the site of infection, than B6 mice, suggesting that local production of anti-MHV antibody contributed to this absence of CTL escape. Additionally, administration of anti-MHV neutralizing antibody to infected B6 mice suppressed the development of CTL escape mutants. These findings indicate a key role for the anti-MHV antibody response in suppressing virus replication, thereby minimizing the emergence and competitive advantage of CTL escape mutants.
PPPYEPTAP Motif Is the Late Domain of Human T-Cell Leukemia Virus Type 1 Gag and Mediates Its Functional Interaction with Cellular Proteins Nedd4 and Tsg101
Authors: Bouamr Fadila , Melillo Jessica A. , Wang Margaret Q. , Nagashima Kunio , de Los Santos Martha , Rein Alan , Goff Stephen P.
Published: October 2003

The human T-cell leukemia virus type 1 (HTLV-1) Gag polyprotein contains two adjacent proline-rich motifs (sequence
PPPY
E
PTAP
) in the C terminus of the matrix domain. Proline-to-alanine mutations were introduced into either or both motifs of HTLV-1 to determine the effect on the release of HTLV-1 virus-like particles from 293T cells. The release of both single mutants was significantly reduced, whereas a double mutation in both motifs abolished the release of the HTLV-1 particles. Two-hybrid and in vitro binding assays showed that the HTLV-1 Gag polyprotein binds both Tsg101 and Nedd4 proteins. The interaction with HTLV-1 Gag required the central WW domain of Nedd4 and the ubiquitin enzyme variant (UEV) domain of Tsg101. We expressed various fragments of Nedd4 and Tsg101 proteins in 293T cells and tested for their ability to interfere with virion release mediated by the HTLV-1 Gag-Pro polyprotein. Fragments consisting of the N-terminal UEV domain of Tsg101 and the central WW and C-terminal Hect domains of Nedd4 protein all caused transdominant inhibition of HTLV-1 particle release. Similarly, inhibition of the proteasome significantly decreased HTLV-1 particle release. Furthermore, the WW domain overexpression caused an early arrest of HTLV-1 particle morphogenesis before the membrane is deformed into the typical half-shell structure. This result suggests that Nedd4 is involved early in budding of HTLV-1.
Resistance of Human Hepatitis Delta Virus RNAs to Dicer Activity
Authors: Chang Jinhong , Provost Patrick , Taylor John M.
Published: October 2003

The endonuclease dicer cleaves RNAs that are 100% double stranded and certain RNAs with extensive but <100% pairing to release ∼21-nucleotide (nt) fragments. Circular 1,679-nt genomic and antigenomic RNAs of human hepatitis delta virus (HDV) can fold into a rod-like structure with 74% pairing. However, during HDV replication in hepatocytes of human, woodchuck, and mouse origin, no ∼21-nt RNAs were detected. Likewise, in vitro, purified recombinant dicer gave <0.2% cleavage of unit-length HDV RNAs. Similarly, rod-like RNAs of potato spindle tuber viroid (PSTVd) and avocado sunblotch viroid (ASBVd) were only 0.5% cleaved. Furthermore, when a 66-nt hairpin RNA with 79% pairing, the putative precursor to miR-122, which is an abundant liver micro-RNA, replaced one end of HDV genomic RNA, it was poorly cleaved, both in vivo and in vitro. In contrast, this 66-nt hairpin, in the absence of appended HDV sequences, was >80% cleaved in vitro. Other 66-nt hairpins derived from one end of genomic HDV, PSTVd, or ASBVd RNAs were also cleaved. Apparently, for unit-length RNAs of HDV, PSTVd, and ASBVd, it is the extended structure with <100% base pairing that confers significant resistance to dicer action.
Construction, Rescue, and Characterization of Vectors Derived from Ovine Atadenovirus
Authors: Löser Peter , Hofmann Christian , Both Gerald W. , Uckert Wolfgang , Hillgenberg Moritz
Published: October 2003

Gene transfer vectors derived from ovine atadenovirus type 7 (OAdV) can efficiently infect a variety of mammalian cells in vitro and in vivo to deliver and express transgenes. However, early OAdV vectors were designed on human mastadenovirus principles prior to the complete characterization of OAdV genes and transcripts. The distinctive arrangement of the OAdV genome has suggested ways to improve OAdV vector design and utility. We therefore developed a cosmid-based approach that allows efficient construction of recombinant ovine atadenovirus genomes in which the transgene is inserted at one of three sites. Viruses were rescued by transfection of viral DNA into a new ovine fetal skin fibroblast producer cell line, HVO156. The suitability of the three insertion sites was compared with respect to virus rescue efficiency, gene expression levels, and genetic stability of the vectors. We found that one vector with a transgene inserted at site 1, between the pVIII and fiber genes, was unstable. Only one vector that carried a transgene at site 2, near the right end of the genome, together with a nearby deletion was rescued. In contrast, several vectors with different transgenes inserted in site 3, between the E4 and RH transcription units, were repeatedly rescued, and these vectors were stable over at least four passages. Transgene orientation in site 3 had only little effect on expression. Finally, a vector carrying a human factor IX cDNA at site 3, when administered intravenously, produced nearly physiological levels of human factor IX in mice. The availability of an efficient method for vector construction and the identification of a new insertion site for virus rescue and gene expression substantially enhance the utility of the OAdV vector system.
Induction of Caspase-Dependent Apoptosis in Cultured Rat Oligodendrocytes by Murine Coronavirus Is Mediated during Cell Entry and Does Not Require Virus Replication
Authors: Liu Yin , Cai Yingyun , Zhang Xuming
Published: October 2003

Murine coronavirus mouse hepatitis virus (MHV) causes demyelination of the central nervous system (CNS) in rats and mice. Apoptotic oligodendrocytes have been detected in the vicinity of the CNS demyelinating lesions in these animals. However, whether MHV can directly induce oligodendrocyte apoptosis has not been documented. Here, we established a rat oligodendrocyte culture that is morphologically and phenotypically indistinguishable from the primary rat oligodendrocytes. Using this culture, we showed that mature rat oligodendrocytes were permissive to MHV infection but did not support productive virus replication. Significantly, oligodendrocytes infected with both live and ultraviolet light-inactivated viruses underwent apoptosis to a similar extent, which was readily detectable at 24 h postinfection as revealed by apoptotic bodies and DNA fragmentation, indicating that MHV-induced apoptosis is mediated during the early stages of the virus life cycle and does not require virus replication. Prior treatment of cells with the lysosomotropic agents NH4Cl and chloroquine as well as the vacuolar proton pump-ATPase inhibitor bafilomycin A1, all of which block the acidification of the endosome, prevented oligodendrocytes from succumbing to apoptosis induced by MHV mutant OBLV60, which enters cells via endocytosis, indicating that fusion between the viral envelope and cell membranes triggers the apoptotic cascade. Treatment with the pan-caspase inhibitor Z-VAD-fmk blocked MHV-induced apoptosis, suggesting an involvement of the caspase-dependent pathway. Our results, thus, for the first time provide unequivocal evidence that infection of oligodendrocytes with MHV directly results in apoptosis. This finding provides an explanation for the destruction of oligodendrocytes and the damage of myelin sheath in MHV-infected CNS and suggests that oligodendrocyte apoptosis may be one of the underlying mechanisms for the pathogenesis of MHV-induced demyelinating diseases in animals.
G541R within the 4070A TM Protein Regulates Fusion in Murine Leukemia Viruses
Authors: O'Reilly Lucille , Roth Monica J.
Published: October 2003

The mutation G541R within the ectodomain of TM was isolated in three independent chimeric enveloped murine leukemia virus (MuLV) viral populations originally impaired in viral passage and in wild-type 4070A. Isolation of G541R in multiple populations suggested it played a critical role in viral envelope function. Using a viral vector system, the observed effects of the G541R mutation within MuLV envelope proteins were pleiotropic and included effects on the regulation of SU-TM interactions and membrane fusion. G541R suppresses enhanced cell-cell fusion events attributable to the absence of the R-peptide yet does not adversely affect virus titers. The ability to suppress cell-cell fusion is dependent on the presence of the C terminus of the amphotropic 4070A SU protein. Within the wild-type 4070A envelope background, the mutation results in a decreased level of Env at the cell surface that is mirrored in the virion. The TM mutation alters recognition of the SU C terminus by a monoclonal antibody, suggestive of an altered conformation. The presence of G541R allowed the virus to achieve a balance between cytopathogenicity and replication and restored productive viral entry.
Mechanism of RNA Recombination in Carmo- and Tombusviruses: Evidence for Template Switching by the RNA-Dependent RNA Polymerase In Vitro
Authors: Cheng Chi-Ping , Nagy Peter D.
Published: October 2003

RNA recombination occurs frequently during replication of tombusviruses and carmoviruses, which are related small plus-sense RNA viruses of plants. The most common recombinants generated by these viruses are either defective interfering (DI) RNAs or chimeric satellite RNAs, which are thought to be generated by template switching of the viral RNA-dependent RNA polymerase (RdRp) during the viral replication process. To test if RNA recombination is mediated by the viral RdRp, we used either a purified recombinant RdRp of
Turnip crinkle carmovirus
or a partially purified RdRp preparation of
Cucumber necrosis tombusvirus
. We demonstrated that these RdRp preparations generated RNA recombinants in vitro. The RdRp-driven template switching events occurred between either identical templates or two different RNA templates. The template containing a replication enhancer recombined more efficiently than templates containing artificial sequences. We also observed that AU-rich sequences promote recombination more efficiently than GC-rich sequences. Cloning and sequencing of the generated recombinants revealed that the junction sites were located frequently at the ends of the templates (end-to-end template switching). We also found several recombinants that were generated by template switching involving internal positions in the RNA templates. In contrast, RNA ligation-based RNA recombination was not detected in vitro. Demonstration of the ability of carmo- and tombusvirus RdRps to switch RNA templates in vitro supports the copy-choice models of RNA recombination and DI RNA formation for these viruses.
Vaccine-Induced Immunopathology during Bovine Respiratory Syncytial Virus Infection: Exploring the Parameters of Pathogenesis
Authors: Antonis Adriaan F. G. , Schrijver Remco S. , Daus Franz , Steverink Paul J. G. M. , Stockhofe Norbert , Hensen Evert J. , Langedijk Johannes P. M. , van der Most Robbert G.
Published: October 2003

The bovine and human respiratory syncytial viruses cause severe lower respiratory tract infections. Effective vaccines against the respiratory syncytial viruses have been lacking since vaccine failures in the 1960s and 1970s. In this report, we describe a bovine respiratory syncytial virus (bRSV) challenge model in which both classical bRSV respiratory infection and vaccine-enhanced immune pathology were reproduced. The classical, formalin-inactivated (FI) bRSV vaccine that has been associated with vaccine failure was efficient in inducing high antibody titers and reducing viral loads but also primed calves for a far more serious enhanced respiratory disease after a bRSV challenge, thereby mimicking the enhanced clinical situation in FI human RSV (hRSV)-immunized and hRSV-infected infants in the 1960s. We show that immunization with FI-bRSV mainly primes a Th2-like inflammatory response that is characterized by a significant eosinophilic influx in the bronchial alveolar lung fluid and lung tissues and high levels of immunoglobulin E serum antibodies. The current model may be useful in the evaluation of new bRSV candidate vaccines for potency and safety.