bluuebunny/crossref_metadata_2025 huggingface dataset:
FACTORS INFLUENCING LEGISLATORS’USE OF POLICY INFORMATION AND IMPLICATIONS FOR PROMOTING GREATER USE
Authors: Webber David J.
Published: June 2005

Despite a tremendous growth in the volume of policy analysis in the past decade, recent research indicates governmental decisionmakers make little direct use of such research; at best, policy‐oriented research alters a decisionmaker's conceptualization of the policy problem over the long term. Previous efforts to explain policymakers’limited use of policy research have generally not considered the opportunities and constraints presented by specific decisionmaking contexts. To a large extent, the research utilization literature has focused on attitudinal and demographic characteristics of decisionmakers and has adopted as a central focus the “two‐communities” theory which asserts that frequent, direct use of policy research by policymakers is unlikely because of the competing worldview and belief systems of policy researchers and policymakers. While the two‐communities theory may partially explain the limited use of policy research, it ignores the institutional incentives motivating decisionmakers. More specifically, this earlier research does not consider the different motivations of policymakers and their relative impact on use of policy information. To expand the explanation of decisionmakers’use of policy information, I postulate a model of legislators’use of policy information that includes four types of factors that influence legislators’propensity to use policy information. These factors‐worldview and attitude toward social science, perception of the way policy issues are considered in the legislature, perception of the legislative structure, and legislative orientations‐combine two‐communities type of independent variables and perceptions of the legislative environment. In this way, policymakers’use of policy information can be examined more realistically and comprehensively.
ECONOMIC MOTIVES FOR “SOCIAL” PROGRAMS: THE NEW FEDERAL DIRECTION
Authors: Jensen J. Martin
Published: June 2005

The Job Training Partnership Act (JTPA), passed by Congress in 1982, is significant federal employment and training legislation for a number of reasons. Most noticeably, it substantially enhanced private sector and state government roles in the administration of such programs. In order to understand both the programmatic impact of JTPA and its likely consequences on subsequent federal employment and training initiatives, it is necessary to look at the politics of the legislation‐especially those interests that are strongly represented through its implementation and those that are not. By doing this, one obtains a better sense of the dimensions of conflict around future employment and training legislation. General guidelines within which Congress should act in subsequent legislative activity are laid out.
FEDERAL EMPLOYMENT AND TRAINING POLICY: A VIEW FROM THE LOCAL LEVEL OF GOVERNMENT
Authors: Pines Marion
Published: June 2005

To maximize the use of human capital, employment and training resources need to be redirected to meet four objectives: (1) reduce welfare dependency by helping people gain skills for self‐sufficiency, (2) prepare a new workforce through better education of youth, (3) retrain dislocated workers, and (4) develop an employment system that links workers and jobs through the coordination of institutions. Suggestions for federal, state, and local initiatives to meet these objectives are spelled out. Particular emphasis is placed upon both a significant revitalization of the federal role in employment and training and a reassessment of traditional approaches.
TURBULENT TRANSITION AND ORGANIZATIONAL CHANCE: RELATING POLICY OUTCOMES TO STRATEGIC ADMINISTRATIVE CAPACITIES
Authors: Boschken Herman L.
Published: June 2005

Much of the literature on public policy with its institutional and legislative emphases conveys the view that organizational effectiveness is primarily dependent on actors external to the agency. That is, laws prescribe programs and operating procedures; overlapping jurisdictions restrict innovative behavior; and the politics o f fund budgeting encourage complacency and incrementalism. This article proposes an alternative set of propositions explaining policy outcomes and agency performance as a function of internal administrative considerations. The propositions are empirically evaluated using data from Pacific Coast port authorities during the “container revolution” and environmental movement. The results suggest that variance in strategic performance can be attributed to (1) perceptions about intergovernmental relations and performance gap, (2) an agency micro‐structure for strategic planning and policy analysis, and (3) a set of intervening administrative variables.
THE PLURALITY OF FACTORS INFLUENCING POLICYMAKINC: SCHOOL REFORM LEGISLATION IN THE AMERICAN STATES, 1982–84
Authors: Shinn Doh C. , Van Der Slik Jack R.
Published: June 2005

This paper attempts to explore recent efforts of American state legislatures to improve the quality of public schools. Which state legislatures have passed school reform legislation? At what aspects of public education is this reform legislation aimed? Are there significant variations across states in such reform legislation? What has contributed most to the passage of such legislation? Is it the socioeconomic characteristics of individual states or the structural and procedural characteristics of their legislative branch that better explain school reform legislation? Analysis of data from the 50 states reveals greater variation in legislative results that reform public schools. The regional locus of most reform was the south. Most importantly, school reform legislation was found to be influenced by the interaction of a multitude of divergent forces. This finding suggests that researchers interested in the determinants of legislative action must move beyond analysis of whether economic or political forces shape legislative outcomes to inquire how economic, political and other forces interact when a given reform effort reaches the legislative halls of the states.
THE USES OF JAIL CONFINEMENT IN THREE COUNTIES
Authors: Jackson Patrick G.
Published: June 2005

This study uses a longitudinal approach to study the characteristics of pretrial inmates and court dispositions of their charges in three jails. Over 1,300 inmates were tracked from the point of booking through court disposition. The results indicate that a majority of the inmates booked into the jails are charged with relatively minor offenses and that most have less serious misdemeanor arrest and conviction histories. Very few or none of the defendants at each site were sentenced to prison and, at most, about one‐fifth of the defendants were sentenced to any additional jail time upon conviction. Policy implications of the results are briefly discussed.
DOING TIME IN THE NEW GENERATION JAIL: INMATE PERCEPTIONS OF GAINS AND LOSSES
Authors: Zupan Linda L. , Stohr‐Gillmore Mary K.
Published: June 2005

Observations of New Generation, podularldirect supervision jail operations indicate that the innovative architectural design and inmate management style provide a safer, more humane environment for inmates while concurrently reducing opportunities for inmates to illegitimately fulfill their needs. In the first part of this paper, we demonstrate how differences in New Generation and traditional jail architecture and operations influence the legitimate and illegitimate means by which inmates seek to fulfill their needs. Using data collected from New Generation and traditional jails, we then compare inmate perceptions of need fulfillment, evaluations of the environment and levels of stress.
A CRITICAL ASSESSMENT OF ELECTRONIC MONITORING IN CORRECTIONS
Authors: Clear Todd R.
Published: June 2005

Electronic monitoring is a recent advance in information gathering for correctional program administration. To be effective in achieving correctional goals, electronic monitors must be applied to offenders when there is some risk of performance problem. Advocates of electronic monitoring argue they can promote public safety, save tax dollars, and enhance correctional performance. Because corrections systems find it so difficult to innovate with offenders who represent a risk to society, these claims are overstated. Electronic monitoring raises short‐term concerns concerning damage to correctional program credibility and long‐term concerns about the enhancement of social control.
THE AMBITIONS OF POLICY DESIGN
Authors: Dryzek John S. , Ripley Brian
Published: June 2005

There is no shortage of warnings concerning the hazards of excessive ambition in consciously‐pursued public policy. In the light of these cautions, this paper considers the appropriate ambitions for policy design. The critics have missed the target. There is no need to fear ambition in policy design, provided that openness in communications about policy is secured.
RURAL AND URBAN POVERTY: AN INCOME/NET WORTH APPROACH
Authors: Lerman Donald L. , Mikesell James J.
Published: June 2005

This paper examines rural and urban changes in the distribution of poverty that would result from modifying the conventional poverty measure to include the annuity value of household net worth.

Use of this new income/wealth measure caused numerous shifts in the location and demography of the poverty population. Among those more often found to be in poverty under the new measure were young, renter, and large central city resident households. Age, homeownership, farm employment, education, retirement status, public assistance participation, and residence in the West were important factors in explaining the divergence of the WH and INC measures. The age and retirement impacts were significantly different in rural and urban areas. Rural residence itself was not an important factor in explaining WH and INC differences.
SOCIAL SCIENCE DISCIPLINES AND POLICY RESEARCH: THE CASE OF POLITICAL SCIENCE
Authors: Rogers James M.
Published: June 2005

It has been argued that social science disciplines influence their members policy research via theoretical focus, methods, norms, and system maintenance mechanisms and that these forces inhibit the usefulness of policy research for policy‐making. Political science is found to influence substantially its members policy research output and to decrease its policy usefulness, primarily by promoting explanations of policy, although the extent of influence and lack of usefulness are less than studies of other disciplines suggest. Whereas highly useful outcome analyses are produced less frequently than many advocates of policy research would hope, a sub‐stantial body of policy research undertakes objectives that when satisfied, particularly in the area of problem definition, provide moderately useful output to decision makers. In addition, policy research output is remarkably diverse substantively, but less so in terms of the purposes it serves.
A REGIME FRAMEWORK FOR IMPLEMENTATION ANALYSIS: COOPERATION AND RECONCILIATION OF FEDERALIST IMPERATIVES
Authors: Stoker Robert P.
Published: June 2005

The diffusion of authority characteristic of federal governance challenges national leadership and complicates even the implementation of policy that promises mutual gains. To explore the problems of implementing national policy in a federalist context, this paper proposes a new approach to analysis, t h e implementation regime framework. From the regime perspective, the essential task of the implementation process is to create a context in which implementors are likely to cooperate to achieve policy goals despite the absence of dominating authority. Drawing upon the literature on international cooperation, the regime framework outlines the contextual conditions and elements of policy design that are likely to lead to cooperation in collective decision making.
THE POLITICS OF LEGISLATIVE OVERSIGHT: MONITORING EDUCATIONAL REFORM IN SIX STATES
Authors: Wohlstetter Priscilla
Published: June 2005

Legislative oversight traditionally has been viewed as a rational tool for achieving democratic accountability. Legislators conduct oversight to ensure consistency between implementation and official policy directives. The major theme of this paper is t h a t oversight is best understood as an integral part of legislative‐administrative politics. By examining how six states monitored recent educational reforms, the author finds that legislators conducted oversight in ways that minimized time commitments and maximized political benefits.

The paper concludes that legislative interest in controlling implementation often was secondary to political considerations. Oversight, therefore, may not really be effective as an accountability tool.
PROBLEM DEFINITION AND THE ALASKA NATIVES: ETHNIC IDENTITY AND POLICY FORMATION
Authors: Korsmo Fae L.
Published: June 2005

This case study of an ethnoterritorial minority's role in agenda setting argues that the extant agenda setting literature does not take into account different cultural perceptions of the definitions of problems. Using the hearings on the Alaska Native Claims Settlement Act of 1971, the differen‐ ces in understanding of the problem by the Alaskan natives who testified before the Committee and the Senators who received the testimony are examined.
Teaching and learning in dental student clinical practice
Authors: Fugill M.
Published: June 2005

Abstract  Clinical learning in restorative dentistry is principally centred around the provision of patient care, yet we know very little about the learning processes occurring within the clinical environment. A study of undergraduate dental student clinical practice used a combination of group interview and questionnaire techniques to explore some of the characteristics of student/teacher interaction that students finds significant, and which they consider to affect their learning of clinical skills. Study data, when analysed, revealed three major categories of teacher or student behaviour which appear to be of importance to students. This paper focuses on one of these, describing a number of behaviours, grouped together under the category of ‘teaching/learning behaviours’. The aim of the paper is to report these results and to discuss their application to clinical teaching of restorative dentistry.
New Claims about Executions and General Deterrence: Déjà Vu All Over Again?
Authors: Berk Richard
Published: June 2005

A number of papers have recently appeared claiming to show that in the United States executions deter serious crime. There are many statistical problems with the data analyses reported. This article addresses the problem of “influence,” which occurs when a very small and atypical fraction of the data dominate the statistical results. The number of executions by state and year is the key explanatory variable, and most states in most years execute no one. A very few states in particular years execute more than five individuals. Such values represent about 1 percent of the available observations. Reanalyses of the existing data are presented showing that claims of deterrence are a statistical artifact of this anomalous 1 percent.
Chronic Care Management for the Elderly: An Opportunity for Gerontological Nurse Practitioners
Authors: Hendrix Cristina C. , Wojciechowski Colleen W.
Published: June 2005

Purpose

To discuss the role that gerontological nurse practitioners (GNPs) may play in providing chronic care management for the elderly.

Data sources

Review of recent literature on chronic care management, personal experience of the authors in caring for older adults under the chronic care management model, and a case study.

Conclusions

GNPs are the most appropriate practitioners to provide and coordinate chronic care management to the population that needs it most—the elderly.

Implications for practice

Demographic shifts in the United States have increased the number of people with chronic illnesses; however, the nation's healthcare delivery system has not significantly evolved to meet the changing needs of its population. As a result, many people, especially older adults, suffer needlessly, and healthcare costs continue to rise. Chronic care management may alleviate older adults’ chronic health problems, reduce expenditures for their health care, and promote their satisfaction and quality of life.
Vertebroplasty and Kyphoplasty for Treatment of Painful Osteoporotic Compression Fractures
Authors: Lemke Denise M.
Published: June 2005

Purpose

To review the pathophysiology of osteoporosis and describe vertebroplasty and kyphoplasty, which are minimally invasive procedures to treat the pain associated with vertebral compression fractures (VCFs).

Data sources

Extensive literature review of osteoporosis, vertebroplasty, and kyphoplasty supplemented by case study and clinical experience in the minimally invasive interventional neuroradiology interventions.

Conclusions

Osteoporosis is a progressive debilitating process that destroys the cancellous bone, weakening the overall integrity and stability of the bone. The loss of bone mass places the individual at increased risk for vertebral body, hip, and wrist fractures. In the past, there was no treatment option to repair vertebral body deformity or instability after osteoporotic VCFs. Management solely relied on the use of nonsteroidal anti‐inflammatory drugs, narcotics, muscle relaxants, and/or orthotic bracing to provide pain relief. VCFs alter the stability of the vertebral body and column, and the lack of stabilization can lead to chronic pain syndrome, immobility, pulmonary compromise, progression of spinal deformity, increase in the risk for additional VCFs, and increase in the risk for comorbidities and mortality related to immobility.

Implications for Practice

Vertebroplasty and kyphoplasty are minimally invasive procedures aimed at pain control, stabilization of the vertebral body, and with kyphoplasty, the ability to provide some correction of deformity with partial restoration of vertebral body height. Providing pain control and stabilization of the vertebral column improves mobility, thus decreasing the potential risks associated with immobility.
Chronic Blood Transfusion Therapy Practices to Treat Strokes in Children with Sickle Cell Disease
Authors: Lindsey Terianne , Watts‐Tate Nutrena , Southwood Elaine , Routhieaux Julie , Beatty Janice , Calamaras Diane , Phillips Melissa , Lea Gwendolyn , Brown Evelyn , DeBaun Michael R.
Published: June 2005

Purpose

To identify variations in practices used by nurses for pediatric patients with sickle cell disease (SCD) receiving chronic blood transfusion therapy for strokes.

Data sources

Descriptive study of a convenience sample of 11 nurses who care for children with SCD from nine institutions completed a closed‐ended questionnaire consisting of 37 items. Responses reflected practice experience with a total of 189 transfused patients with SCD.

Conclusions

A wide range of nursing practices exists for blood transfusion therapy for children with SCD and strokes. Manual partial exchange transfusion (66%) was the most commonly used method for blood transfusion in children with strokes reported among the nurses surveyed. Simple transfusions and erythrocytapheresis account for 21% and 13% of the practices reported. Opportunities exist to establish evidence‐based nursing care guidelines to improve the care of children with strokes receiving blood transfusion therapy.

Implications for practice

A wide range of local standard care guidelines for blood transfusion therapy exists. The results of this survey indicate that partial manual exchange transfusion is the most commonly used method of chronic blood transfusion therapy in children with SCD and stroke despite the fact that the magnitude of benefit in comparison with simple transfusion has not been established. Factors such as peripheral venous access, compliance with current chelation regimen, and the presence of antibodies are important considerations in the choice of method.
Tyrosine Phosphorylation of the Chlamydial Effector Protein Tarp Is Species Specific and Not Required for Recruitment of Actin
Authors: Clifton Dawn R. , Dooley Cheryl A. , Grieshaber Scott S. , Carabeo Reynaldo A. , Fields Kenneth A. , Hackstadt Ted
Published: June 2005

Chlamydiae are obligate intracellular pathogens that efficiently induce their endocytosis by susceptible eukaryotic host cells. Recently, a
Chlamydia trachomatis
type III secreted effector protein, Tarp, was found to be translocated and tyrosine phosphorylated at the site of entry and associated with the recruitment of actin that coincides with endocytosis.
C. trachomatis
Tarp possesses up to six direct repeats of approximately 50 amino acids each. The majority of the tyrosine residues are found within this repeat region. Here we have ectopically expressed distinct domains of Tarp in HeLa 229 cells and demonstrated that tyrosine phosphorylation occurs primarily within the repeat region, while recruitment of actin is mediated by the C-terminal domain of the protein. A comparison of other sequenced chlamydial genomes revealed that each contains an ortholog of Tarp, although
Chlamydia muridarum
,
Chlamydophila caviae
, and
Chlamydophila pneumoniae
Tarp lack the large repeat region. Immunofluorescence and immunoblotting using an antiphosphotyrosine antibody show no evidence of phosphotyrosine at the site of entry of
C. muridarum
,
C. caviae
, and
C. pneumoniae
, although each species similarly recruits actin. Ectopic expression of full-length
C. trachomatis
and
C. caviae
Tarp confirmed that both recruit actin but only
C. trachomatis
Tarp is tyrosine phosphorylated. The data indicate that the C-terminal domain of Tarp is essential for actin recruitment and that tyrosine phosphorylation may not be an absolute requirement for actin recruitment. The results further suggest the potential for additional, unknown signal transduction pathways associated specifically with
C. trachomatis
.
Noninvasive Biophotonic Imaging for Monitoring of Catheter-Associated Urinary Tract Infections and Therapy in Mice
Authors: Kadurugamuwa Jagath L. , Modi Kshitij , Yu Jun , Francis Kevin P. , Purchio Tony , Contag Pamela R.
Published: June 2005

Urinary tract infections (UTIs) are among the most common bacterial infections acquired by humans, particularly in catheterized patients. A major problem with catheterization is the formation of bacterial biofilms on catheter material and the risk of developing persistent UTIs that are difficult to monitor and eradicate. To better understand the course of UTIs and allow more accurate studies of in vivo antibiotic efficacy, we developed a catheter-based biofilm infection model with mice, using bioluminescently engineered bacteria. Two important urinary tract pathogens,
Pseudomonas aeruginosa
and
Proteus mirabilis
, were made bioluminescent by stable insertion of a complete
lux
operon. Segments of catheter material (precolonized or postimplant infected) with either pathogen were placed transurethrally in the lumen of the bladder by using a metal stylet without surgical manipulation. The bioluminescent strains were sufficiently bright to be readily monitored from the outside of infected animals, using a low-light optical imaging system, including the ability to trace the ascending pattern of light-emitting bacteria through ureters to the kidneys. Placement of the catheter in the bladder not only resulted in the development of strong cystitis that persisted significantly longer than in mice challenged with bacterial suspensions alone but also required prolonged antibiotic treatment to reduce the level of infection. Treatment of infected mice for 4 days with ciprofloxacin at 30 mg/kg of body weight twice a day cured cystitis and renal infection in noncatheterized mice. Similarly, ciprofloxacin reduced the bacterial burden to undetectable levels in catheterized mice but did not inhibit rebound of the infection upon cessation of antibiotic therapy. This methodology easily allows spatial information to be monitored sequentially throughout the entire disease process, including ascending UTI, treatment efficacy, and relapse, all without exogenous sampling, which is not possible with conventional methods.
Development of Secondary Inclusions in Cells Infected byChlamydia trachomatis
Authors: Suchland Robert J. , Rockey Daniel D. , Weeks Sara K. , Alzhanov Damir T. , Stamm Walter E.
Published: June 2005

The chlamydiae are obligate intracellular bacteria that occupy a nonacidified vacuole (the inclusion) during their entire developmental cycle. These bacteria produce a set of proteins (Inc proteins) that localize to the surface of the inclusion within infected cells.Chlamydia trachomatisIncA is also commonly found in long fibers that extend away from the inclusion. We used standard and confocal immunofluorescence microscopy to demonstrate that these fibers extend to newly developed inclusions, termed secondary inclusions, within infected cells. Secondary inclusions observed at early time points postinfection were devoid of chlamydial reticulate bodies. Later in the developmental cycle, secondary inclusions containing variable numbers of reticulate bodies were common. Reticulate bodies were also observed within the IncA-laden fibers connecting primary and secondary inclusions. Quantitative differences in secondary inclusion formation were found among clinical isolates, and these differences were associated with serovar. Isolates of serovar G consistently produced secondary inclusions at the highest frequency (P< 0.0001). Similar quantitative studies demonstrated that secondary inclusion formation was associated with segregation of inclusions to daughter cells following cytokinesis. We conclude that the production of secondary inclusions via IncA-laden fibers allows chlamydiae to generate an expanded intracellular niche in which they can grow and may provide a means for continuous infection within progeny cells following cell division.
Posttranslational Modification of RecombinantPlasmodium falciparumApical Membrane Antigen 1: Impact on Functional Immune Responses to a Malaria Vaccine Candidate
Authors: Giersing Birgitte , Miura Kazutoyo , Shimp Richard , Wang Jin , Zhou Hong , Orcutt Andrew , Stowers Anthony , Saul Allan , Miller Louis H. , Long Carole , Singh Sanjay
Published: June 2005

Recombinant apical membrane antigen 1 (AMA1) is a leading vaccine candidate forPlasmodium falciparummalaria, as antibodies against recombinantP. falciparumAMA1 (PfAMA1) interrupt merozoite invasion into erythrocytes. In order to investigate the role of posttranslational modification in modulating the functional immune response to recombinant AMA1, two separate alleles of PfAMA1 (FVO and 3D7), in which native N-glycosylation sites have been mutated, were produced usingEscherichia coliand aPichia pastorisexpression system. RecombinantPichia pastorisAMA1-FVO (PpAMA1-FVO) and PpAMA1-3D7 are O-linked glycosylated, and 45% of PpAMA1-3D7 is nicked, though all four recombinant molecules react with conformation-specific monoclonal antibodies. To address the immunological effect of O-linked glycosylation, we compared the immunogenicity ofE. coliAMA1-FVO (EcAMA1-FVO) and PpAMA1-FVO antigens, since both molecules are intact. The effect of antigen nicking was then investigated by comparing the immunogenicity of EcAMA1-3D7 and PpAMA1-3D7. Our data demonstrate that there is no significant difference in the rabbit antibody titer elicited towards EcAMA1-FVO and PpAMA1-FVO or to EcAMA1-3D7 and PpAMA1-3D7. Furthermore, we have demonstrated that recombinant AMA1 (FVO or 3D7), whether expressed and refolded fromE. colior produced from thePichiaexpression system, is equivalent and mimics the functionality of the native protein in in vitro growth inhibition assay experiments. We conclude that in the case of recombinant AMA1, theE. coli- andP. pastoris-derived antigens are immunologically and functionally equivalent and are unaffected by the posttranslational modification resulting from expression in these two systems.
Characterization of Native Outer Membrane Vesicles fromlpxLMutant Strains ofNeisseria meningitidisfor Use in Parenteral Vaccination
Authors: Fisseha Makda , Chen Ping , Brandt Brenda , Kijek Todd , Moran Elizabeth , Zollinger Wendell
Published: June 2005

Native outer membrane vesicles (NOMV) ofNeisseria meningitidisconsist of intact outer membrane and contain outer membrane proteins (OMP) and lipooligosaccharides (LOS) in their natural conformation and membrane environment. NOMV have been safely used intranasally in P1 studies with encouraging results, but they are too toxic for parenteral vaccination. We now report the preparation and characterization oflpxLmutants that express LOS with reduced toxicity, and the evaluation of the potential of NOMV from these strains for use as a parenteral vaccine. A series of deletion mutants were prepared with knockouts of one or more of thelpxL1,lpxL2, orsynXgenes. The ΔlpxL2mutants had a reduced growth rate, reduced level of LOS expression, and increased sensitivity to surfactants. In addition, ΔsynXΔlpxL2double mutants had reduced viability in stationary phase. The ΔlpxL1ΔlpxL2double mutant behaved essentially the same as the ΔlpxL2single mutant. LOS from bothlpxLmutant strains exhibited altered migration on polyacrylamide gels. The LOS of ΔlpxL2mutants of L3,7 strains were fully sialylated. NOMV prepared fromlpxL2mutants was about 200-fold less active than wild-type NOMV in rabbit pyrogen tests and in tumor necrosis factor alpha release assays. Bactericidal titers induced in animals by ΔlpxL2mutant NOMV were lower than those induced by ΔlpxL1or wild-type NOMV. However, immunogenicity could be largely restored by use of an adjuvant. These results provide evidence that NOMV from ΔlpxL2mutant strains will be safe and immunogenic in humans when given parenterally.
Expression of the
nfa1
Gene Cloned from Pathogenic
Naegleria fowleri
in Nonpathogenic
N. gruberi
Enhances Cytotoxicity against CHO Target Cells In Vitro
Authors: Jeong Seok-Ryoul , Lee Sang-Chul , Song Kyoung-Ju , Park Sun , Kim Kyongmin , Kwon Myung-Hee , Im Kyung-il , Shin Ho-Joon
Published: June 2005

The pathogenic amoeba
Naegleria fowleri
has a 360-bp
nfa1
gene that encodes the Nfa1 protein (13.1 kDa), which is located in the pseudopodia of the amoeba, and an anti-Nfa1 antibody reduces
N. fowleri
-induced mammalian-cell cytotoxicity in vitro. In contrast, an anti-Nfa1 antibody cannot detect Nfa1 protein expression in the nonpathogenic amoeba
Naegleria gruberi
, which also possesses the
nfa1
gene. In the present study, the
nfa1
gene cloned from pathogenic
N. fowleri
was transfected into nonpathogenic
N. gruberi
to determine whether it was related to pathogenicity. The
nfa1
gene was initially inserted into a eukaryotic transfection vector, pEGFP-C2, containing a cytomegalovirus promoter and the green fluorescent protein (GFP) gene, and was designed as pEGFP-C2/
nfa1
UTR (
nfa1
UTR contains 5′ upstream regions, the
nfa1
open reading frame, and 3′ downstream regions). After transfection, the green fluorescence was observed in the cytoplasm of
N. gruberi
trophozoites. These transfectants were preserved for more than 9 months after selection. The transfected
nfa1
gene was observed by PCR using
nfa1
- and vector-specific primers in the genomic DNA of
N. gruberi
transfected with the pEGFP-C2/
nfa1
UTR vector. In addition, the
nfa1
and GFP genes were identified by reverse transcription-PCR in transgenic
N. gruberi
. The Nfa1 protein expressed in transgenic
N. gruberi
was identified as a 13.1-kDa band by Western blotting using an anti-Nfa1 antibody. Finally,
N. gruberi
transfected with the pEGFP-C2/
nfa1
UTR vector was found to have enhanced cytotoxicity against CHO cells compared with naïve
N. gruberi
.
Protective Properties of Rifampin-Resistant Rough Mutants ofBrucella melitensis
Authors: Adone R. , Ciuchini F. , Marianelli C. , Tarantino M. , Pistoia C. , Marcon G. , Petrucci P. , Francia M. , Riccardi G. , Pasquali P.
Published: June 2005

Vaccination againstBrucellainfections in animals is usually performed by administration of live attenuated smoothB. abortusstrain S19 andB. melitensisstrain Rev1. They are proven effective vaccines againstB. abortusin cattle and againstB. melitensisandB. ovisin sheep and goats, respectively. However, both vaccines have the main drawback of inducing O-polysaccharide-specific antibodies that interfere with serologic diagnosis of disease. In addition, they retain residual virulence, being a cause of abortion in pregnant animals and infection in humans. To overcome these problems, one approach is to develop defined rough mutantBrucellastrains lacking O antigen of lipopolysaccharide.B. abortusrough strain RB51, a rifampin-resistant mutant of virulent strainB. abortus2308, is used as a vaccine againstB. abortusinfection in cattle in some countries. However, RB51 is not effective in sheep, and there is only preliminary evidence that it is effective in goats. In this study, we tested the efficacies of six rifampin-resistant rough strains ofB. melitensisin protecting BALB/c mice exposed toB. melitensisinfection. The protective properties, as well as both humoral and cellular immune responses, were assessed in comparison with those provided byB. melitensisRev1 andB. abortusRB51 vaccines. The results indicated that these rough mutants were able to induce a very good level of protection againstB. melitensisinfection, similar to that provided by Rev1 and superior to that of RB51, without inducing antibodies to O antigen. In addition, allB. melitensismutants were able to stimulate good production of gamma interferon. The characteristics of these strains encourage further evaluation of them as alternative vaccines to Rev1 in primary host species.
Identification, Cloning, and Expression of the CAMP-Like Factor Autotransporter Gene (
cfa
) of
Bartonella henselae
Authors: Litwin Christine M. , Johnson Joel M.
Published: June 2005

The CAMP reaction was first described by Christie et al. (R. Christie, N. E. Atkins, and E. Munch-Petersen, Aust. J. Exp. Biol.
22:
197-200, 1944) as the synergistic lysis of sheep red blood cells by
Staphylococcus aureus
sphingomyelinase and CAMP factor (cohemolysin), a secreted protein from group B streptococci. We observed a CAMP-like reaction when
Bartonella henselae
was grown in close proximity to
S. aureus
on 5% sheep blood agar. This study describes the cloning, sequencing, and characterization of a CAMP-like factor autotransporter gene (
cfa
) from
B. henselae
. A cosmid library of
B. henselae
ATCC 49793 was constructed using SuperCos1 in
Escherichia coli
XL1-Blue MR. Cosmids were screened for the CAMP reaction, and a quantitative cohemolysis microtiter assay was developed using purified sphingomyelinase. Cosmid clones with the strongest cohemolytic reaction had similar restriction enzyme patterns. A DNA fragment that expressed the cohemolysin determinant was subcloned in a 7,200-bp StuI-BamHI fragment which contained a 6,024-bp open reading frame. The deduced amino acid sequence showed homology to the family of autotransporters. The autotransporters are a group of proteins that mediate their own export through the outer membrane. They contain an N-terminal passenger region, the α-domain, and a C-terminal transporter region, the β-domain. The α-domain contained four, nearly identical 42-amino-acid repeats and showed homology to the family of RTX (repeat in toxin) hemolysins. The concentrated supernatant of the recombinant strain expressed a protein with a molecular mass of 180 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis consistent with the calculated molecular weight of the secreted α-domain. In conclusion, we have characterized a novel secreted cohemolysin autotransporter protein of
B. henselae
.
Sequence Diversity and Antigenic Variation at the
rag
Locus of
Porphyromonas gingivalis
Authors: Hall Lucinda M. C. , Fawell Stuart C. , Shi Xiaoju , Faray-Kele Marie-Claire , Aduse-Opoku Joseph , Whiley Robert A. , Curtis Michael A.
Published: June 2005

The
rag
locus of
Porphyromonas gingivalis
W50 encodes RagA, a predicted
tonB
-dependent receptor protein, and RagB, a lipoprotein that constitutes an immunodominant outer membrane antigen. The low G+C content of the locus, an association with mobility elements, and an apparent restricted distribution in the species suggested that the locus had arisen by horizontal gene transfer. In the present study, we have demonstrated that there are four divergent alleles of the
rag
locus. The original
rag
allele found in W50 was renamed
rag-1
, while three novel alleles,
rag-2
to
rag-4
, were found in isolates lacking
rag-1
. The three novel alleles encoded variants of RagA with 63 to 71% amino acid identity to RagA1 and each other and variants of RagB with 43 to 56% amino acid identity. The RagA/B proteins have homology to numerous
Bacteroides
proteins, including SusC/D, implicated in polysaccharide uptake. Monoclonal and polyclonal antibodies raised against RagB1 of
P. gingivalis
W50 did not cross-react with proteins from isolates carrying different alleles. In a laboratory collection of 168 isolates, 26% carried
rag-1
, 36% carried
rag-2
, 25% carried
rag-3
, and 14% carried
rag-4
(including the type strain, ATCC 33277). Restriction profiles of the locus in different isolates demonstrated polymorphism within each allele, some of which is accounted for by the presence or absence of insertion sequence elements. By reference to a previously published study on virulence in a mouse model (M. L. Laine and A. J. van Winkelhoff, Oral Microbiol. Immunol.
13:
322-325, 1998), isolates that caused serious disease in mice were significantly more likely to carry
rag-1
than other
rag
alleles.
Chlamydia pneumoniaeAugments the Oxidized Low-Density Lipoprotein-Induced Death of Mouse Macrophages by a Caspase-Independent Pathway
Authors: Yaraei Kambiz , Campbell Lee Ann , Zhu Xiaodong , Liles W. Conrad , Kuo Cho-chou , Rosenfeld Michael E.
Published: June 2005

Chlamydia pneumoniaeis a common respiratory pathogen that is associated with an increased risk of cardiovascular disease. However, the mechanisms by whichC. pneumoniaecontributes to cardiovascular disease have not been determined yet.C. pneumoniaeinfection may accelerate the death of cells within atherosclerotic lesions and contribute to the formation of unstable lesions. To test this hypothesis, the impact ofC. pneumoniaeinfection on the death of lipid-loaded mouse macrophages was investigated. It was observed that RAW 264.7 cells are highly susceptible to the toxic effects of oxidized low-density lipoprotein (LDL) and exhibit markers of cell death within 24 h of treatment with as little as 5 μg/ml oxidized LDL. Subsequent infection with either liveC. pneumoniaeor heat-killed or UV-inactivatedC. pneumoniaeat a low multiplicity of infection for 24 to 72 h stimulated both additional binding of annexin V and the uptake of propidium iodide. Thus,C. pneumoniaeaugments the effects of oxidized LDL on cell death independent of a sustained infection. However, unlike oxidized LDL,C. pneumoniaeinfection does not activate caspase 3 or induce formation of the mitochondrial transition pore or the fragmentation of DNA, all of which are classical markers of apoptosis. Furthermore, primary bone marrow macrophages isolated from mice deficient in Toll-like receptor 2 (TLR-2) but not TLR-4 are resistant toC. pneumoniae-induced death. These data suggest thatC. pneumoniaekills cells by a caspase-independent pathway and that the process is potentially mediated by activation of TLR-2.
Profile of Gene Expression in a Murine Model of Allergic Bronchopulmonary Aspergillosis
Authors: Kurup Viswanath P. , Raju Raghavan , Manickam Pachiappan
Published: June 2005

Allergic bronchopulmonary aspergillosis (ABPA) results from the interactions of theAspergillusallergens and immune system of the patients. We studied the gene expression profile in a mouse model of ABPA. Of the 12,000 genes studied, 1,300 genes showed enhanced expression and represent chemokine, cytokine, growth factor, signal transduction, and transmembrane receptor genes as well as genes related to arginine metabolism.
MATRIX TRACE INEQUALITIES RELATED TO UNCERTAINTY PRINCIPLE
Authors: KOSAKI HIDEKI
Published: June 2005

In their recent article, Luo and Zhang conjectured the matrix trace inequality mentioned in the introduction below, which is motivated by uncertainty principle. We present a proof for the conjectured inequality.
STATIONARY SOLUTIONS OF A SELECTION MUTATION MODEL: THE PURE MUTATION CASE
Authors: CALSINA ÀNGEL , CUADRADO SÍLVIA
Published: June 2005

A selection mutation equations model for the distribution of individuals with respect to the age at maturity is considered. In this model we assume that a mutation, perhaps very small, occurs in every reproduction where the noncompactness of the domain of the structuring variable and the two-dimensionality of the environment are the main features. Existence of stationary solutions is proved using the theory of positive semigroups and the infinite-dimensional version in Banach lattices of the Perron Frobenius theorem. The behavior of these stationary solutions when the mutation is small is studied.
Voltage-Dependent Calcium Currents in Trigeminal Motoneurons of Early Postnatal Rats: Modulation by 5-HT Receptors
Authors: Hsiao Chie-Fang , Wu Nanping , Chandler Scott H.
Published: June 2005

Trigeminal motoneurons relay the final output signals generated within the oral-motor pattern generating circuit(s) to muscles for execution of various motor patterns. In recent years, these motoneurons were shown to possess voltage dependent nonlinear membrane properties that allow them to actively participate in sculpting their final output. A complete understanding of the factors controlling trigeminal motoneuronal (TMN) discharge during oral-motor activity requires, at a minimum, a detailed understanding of the palette of ion channels responsible for membrane excitability and a determination of whether these ion channels are targets for modulation. Toward that end, we studied in detail the properties of calcium channels in TMNs and their susceptibility to modulation by 5-HT in rat brain slices. We found that based on pharmacological and voltage-dependent properties, high-voltage-activated (HVA) N-type [ω-conotoxin GVIA (ω-CgTX)]-sensitive, and to a lesser extent P/Q-type [ω-agatoxin IVA (ω-Aga IVA)]-sensitive, calcium channels make up the majority of the whole cell calcium current. 5-HT (5.0 μM) decreased HVA current by 31.3 ± 2.2%, and the majority of this suppression resulted from reduction of current flow through N- and P/Q-type calcium channels. In contrast, 5-HT had no effect on low-voltage-activated (LVA) current amplitude in TMNs. HVA calcium current inhibition was mimicked by 5-CT, a 5-HT1 receptor agonist, and by R(+)-8-hydroxydipropylaminotetralin hydrobromide (8-OH-DPAT), a specific 5-HT1A agonist. The effects of 5-HT were blocked by the 5-HT1A antagonist 1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl]piperazine hydrobromide (NAN-190) but not by ketanserin, a 5-HT2/1C antagonist. Under current clamp, ω-CgTX and 5-HT were most effective in suppressing the mAHP and both increased the spike frequency and input/output gain in response to current injection. Calcium current modulation by 5-HT1A receptors likely is an important mechanism to fine tune the input/output gain of TMNs in response to small incoming synaptic inputs and accounts for some of the previously reported effects of 5-HT on TMN excitability during tonic and burst activity during oral-motor behavior.
Synaptically Released and Exogenous ACh Activates Different Nicotinic Receptors to Enhance Evoked Glutamatergic Transmission in the Lateral Geniculate Nucleus
Authors: Guo Jian-Zhong , Liu Yingbing , Sorenson Eva M. , Chiappinelli Vincent A.
Published: June 2005

The effects of activation of nicotinic acetylcholine receptors (nAChRs) on glutamatergic transmission in the ventral lateral geniculate nucleus (LGNv) were examined in chick brain slices. Whole cell recordings showed that monosynaptic postsynaptic currents (PSCs) evoked in LGNv neurons by optic tract stimulation were blocked by glutamate receptor antagonists. Exogenously applied nicotine (0.5 μM), choline (1 mM), or acetylcholine (ACh, 100 μM) markedly increased (>3-fold) these evoked PSCs. Potentiation by ACh was dose-dependent and did not desensitize during a 5-min application. In a second set of experiments, the effect of releasing endogenous ACh by stimulating the lateral portion of the LGNv through a separate conditioning electrode before optic tract stimulation was examined. Conditioning stimulation trains increased PSCs by an average of 5.2-fold, an effect dependent on both the intensity and number of conditioning pulses. This increase in PSC amplitude was most likely caused by released ACh activating α6- and/or α3-containing nAChRs because it was blocked by 100 nM α-conotoxin MII, 100 nM dihydro-β-erythroidine (DHβE), and 0.1–1.0 μM methyllycaconitine (MLA). In contrast, exogenously applied ACh increased PSC amplitude by activating a pharmacologically different population of nAChRs because this effect was inhibited by 100 nM α-bungarotoxin, 50 nM MLA, and a high concentration (30 μM) of DHβE, indicating that α7- and/or α8-containing receptors were involved. The results are consistent with a model whereby α6- and/or α3-containing nAChRs on retinal ganglion cell nerve terminals are located preferentially at cholinergic synapses, whereas α7- and/or α8-containing receptors are primarily extrasynaptic.
Nasal Rinsing with Hypertonic Solution: An Adjunctive Treatment for Pediatric Seasonal Allergic Rhinoconjunctivitis
Authors: Garavello Werner , Di Berardino Federica , Romagnoli Marco , Sambataro Giuseppe , Gaini Renato Maria
Published: June 2005

Background: Recent but limited evidence suggests that nasal lavage with hypertonic saline may be useful as an adjunctive treatment modality in the management of pediatric allergic rhinitis. The aim of this study was to clarify whether nasal irrigation with hypertonic solution should be routinely recommended to children with seasonal grass pollen rhinoconjunctivitis. Methods: Fourty-four children with seasonal grass pollen rhinoconjunctivitis were recruited. Twenty-two patients were random ized to receive three-times daily nasal rinsing with hypertonic saline during the pollen season, which lasted 7 weeks. Twenty-two patients were allocated to receive no nasal irrigation and were used as controls. Twenty patients per group completed the study. A mean daily rhinoconjunctivitis score based on the presence of nasal discharge and obstruction as well as ocular symptoms as reddening and itching were calculated for each week of the pollen season. Patients were allowed to use oral antihistamines when required and the mean number of drugs taken per week was also calculated. Results: The mean weekly rhinoconjunctivitis score in the active group was reduced during the whole pollen period. This difference was statistically significant in week 6 and 7 of therapy. A markedly reduced intake of oral antihistamines was also observed in patients allocated to nasal rinsing, being statistically significant in 5 of the 7 weeks. No adverse effect was reported in the active group. Conclusions: This study supports the use of nasal rinsing with hypertonic saline in the pediatric patient with seasonal allergic rhinoconjunctivitis. This treatment proved to be tolerable, inexpensive and effective.