bluuebunny/crossref_metadata_2025 huggingface dataset:
Hormonal factors and respiratory health in women – a review
Authors: Real F. G. , Svanes C. , Macsali F. , Omenaas E. R.
Published: September 2008

Sex hormones appear to play an important role in the lung health of women. This is, however, poorly understood and, in most aspects, poorly investigated; and the literature has been contradictory and confusing. This review presents recent research concerning the involvement of sex hormones in respiratory health of adult women, using the population surveys European Community Respiratory Health Survey and Respiratory Health in Northern Europe.

Respiratory health varied substantially according to hormonal and metabolic conditions. First, menopause was associated with lower lung function and more respiratory symptoms, especially among lean women. Second, hormonal replacement therapy (HRT) was associated with increased risk for asthma and wheeze; also, this association was particularly strong among lean women. Third, women with irregular menstruations in fertile age had more asthma, particularly allergic asthma, and reduced lung function, independently of body mass index (BMI) and physical activity. The findings were consistent across cultural and geographical borders. Our studies revealed that considering interplay between hormonal and metabolic factors is a clue to understand the effects of female sex hormones on the airways. A BMI of around 24–25 kg/m2 appeared to be optimal; women with this BMI had no increase in respiratory health problems when reaching menopause or using HRT, and women in fertile age with this BMI had optimal lung function independently of menstrual status.

In conclusion, female sex hormones appear to play a most important role for lung health in women. Further research on effects of sex hormones on the airways should take into account potential interplay with metabolic factors.

Please cite this paper as: Real FG, Svanes C, Macsali F and Omenaas ER. Hormonal factors and respiratory health in women – a review. The Clinical Respiratory Journal 2008; 2: 111–119.
Concomitant obstructive sleep apnea and chronic obstructive pulmonary disease: study design – the OLIN OSAS‐COPD study
Authors: Larsson L.‐G. , Lindberg A.
Published: September 2008

Background:  Obstructive sleep apnea (OSA) and chronic obstructive pulmonary disease (COPD) are both common disorders. Concomitant disease, overlap syndrome, is reported to cause a more severe condition.

Aim:  To describe a study designed to evaluate the prevalence of overlap syndrome in the general population, and further, to assess the impact of overlap syndrome on cardiovascular morbidity and health‐related quality of life. A secondary aim is to evaluate screening oxygen saturation (Sa02) by pulse oxymetry.

Material and Method:  From the last examination of the OLIN‐studies (Obstructive Lung Disease in Northern Sweden), cohorts I‐IV all subjects with FEV1 ≤ 50 percent predicted or Sa02 ≤ 93 percent predicted were identified (Phase 1). They are invited to a program including lab tests, spirometry, arterial blood gases, chest x‐ray, echocardiography, sleep studies and health‐related quality of life (HRQL) questionnaires. In Phase 2, a random sample of subjects reporting snoring as a problem (n = 100) will be invited to a limited program above all including sleep studies and HRQL questionnaires; and in Phase 3, a random sample (n = 100) will be invited to a similar program.

Summary:  The collection of data in Phase 1 is in progress and will be completed by the end of year 2007. Phase 2 will start year 2008.

Please cite this paper as: Larsson L‐G and Lindberg A. Concomitant obstructive sleep apnea and chronic obstructive pulmonary disease: study design – the OLIN OSAS‐COPD study. The Clinical Respiratory Journal 2008; 2: 120–122.
Genetics of chronic obstructive pulmonary disease: a case–control study in Bergen, Norway
Authors: Sørheim I.‐C. , Gulsvik A.
Published: September 2008

Aims:  Chronic obstructive pulmonary disease (COPD) is a complex disease influenced by multiple genetic and environmental factors. This short communication gives a description of the preliminary genetic results from a case‐control study in Bergen, Norway.

Methods:  A large case‐control study in Bergen in 2003–2005 with 6365 invited subjects generated 1954 cases or controls. The overall attendance of invited subjects was 60%, but causes of non‐attendance varied considerably among different sources of recruitment.

Results:  In this case‐control study, the candidate gene SERPINE2 on chromosome 2q has demonstrated significant association to COPD. However, only weak or lacking associations have so far been observed for the other candidate genes examined.

Conclusion:  These findings provide support for SERPINE2 as a COPD susceptibility gene in the Norwegian population.

Please cite this paper as: Sørheim I‐C and Gulsvik A. Genetics of chronic obstructive pulmonary disease in Bergen, Norway: a case–control study. The Clinical Respiratory Journal 2008; 2: 129–131.
Providing a Dental Home for Pregnant Women: A Community Program to Address Dental Care Access – A Brief Communication
Authors: Milgrom Peter , Ludwig Sharity , Shirtcliff R. Mike , Smolen Darlene , Sutherland Marilynn , Gates Patricia A. , Weinstein Philip
Published: September 2008

Objective: This paper describes a community‐based intervention to provide a dental home for women covered by Medicaid in Klamath County, Oregon. In 2001, 8.8 percent of pregnant women served by Medicaid in Oregon received care. The long‐term goal of the program is to promote preventive oral care for both mothers and their new infants. Methods: Pregnant women received home/Women, Infant and Children visits and were assigned a dental home under a dental managed care program [Dental Care Organization (DCO)]. All initial care was provided at the Oregon Institute of Technology Dental Hygiene Clinic under the contract with the DCO. Emergency, preventive, and restorative care was provided. Results: Between February 2004 and January 2006, 503 pregnant women were identified; 421 women were contactable. Of these, 339 received home visits (339/421, 80.5 percent) and 235 received care (235/339, 69.3 percent). Overall, 55.8 percent of eligible women received care (235/421). Most who did not have a visit either moved or were not the caretaker of the baby. The missed appointment rate was 9 percent. Conclusion: A community health partnership led to a successful and sustainable model extending care to pregnant women and is being extended to promote preventive care for both new mothers and their offspring.
Opinions of Early Head Start Staff about the Provision of Preventive Dental Services by Primary Medical Care Providers
Authors: Mathu‐Muju Kavita R. , Lee Jessica Y. , Zeldin Leslie P. , Rozier R. Gary
Published: September 2008

Objective: This study investigates the opinions of the Early Head Start (EHS) staff about physicians and nurses providing preventive dental services for children in EHS. Methods: A cross‐sectional survey was undertaken of the EHS staff having contact with families in EHS programs in North Carolina (NC). A self‐completed questionnaire solicited their opinions (agree, disagree, don't know) about whether physicians and nurses can “provide preventive dental care” and “identify dental problems” in infants and toddlers. Staff knowledge (four items) and attitudes (five items) were tested for their association with whether staff had an opinion (agree/disagree versus don't know) and if so, what that opinion was (agree versus disagree) using the generalized estimating equation method. Results: Questionnaires were completed by 476 staff (98 percent response) in 18 programs (100 percent response). The majority of staff believed that physicians and nurses can provide preventive dental services (66 percent) and identify dental problems (52 percent). Staff placing importance on ensuring access to dental care and who were knowledgeable about fluoride uses were more likely to have an opinion. Among staff with an opinion, those familiar with the NC program where these services are provided in medical offices were more likely to agree that physicians and nurses can provide preventive services and identify problems. Conclusions: Although the opinions of the majority of the EHS staff are not a barrier to using primary medical care providers to deliver preventive dental care, education is needed for staff who are unfamiliar with this approach.
Follow‐Up Preventive Dental Visits for Medicaid‐Enrolled Children in the Medical Office
Authors: Quiñonez Rocio Beatriz , Pahel Bhavna T. , Rozier R. Gary , Stearns Sally C.
Published: September 2008

Objective: To describe the frequency and determinants of follow‐up preventive oral health visits at medical offices among children screened for dental disease, receiving fluoride varnish and counseling.

Methods: Parents of Medicaid children enrolled in a clinical trial completed a baseline questionnaire before their child's medical visit. The providers completed patient dental encounter forms at each visit, documenting dental services, caries risk, and dental disease. Questionnaires, encounter forms, and Medicaid claims were linked to create a database with information on visits, child and caregiver characteristics, and oral health practices. Descriptive and multivariate analyses assessed associations of variables with likelihood of follow‐up visits.

Results: A total of 744 children with mean age of 15 months at enrollment had 1,415 oral health visits. Children averaged 0.9 follow‐up oral health visits and 1.3 follow‐up well‐child visits. Fewer children had follow‐up oral health visits (55 percent) than well‐child visits (70 percent), but children with a baseline preventive dental visit at a younger age had more visits with shorter intervals. Caregivers reporting greater numbers of children and putting the child to bed with the bottle had more subsequent visits. Older age of child, male child, and caregiver education ≤12 years were associated with fewer follow‐up visits.

Conclusions: Children with preventive dental services in medical offices have similar numbers of oral health and well‐child visits, with both below recommended numbers. Strategies to increase these services may need to be tied to those aimed at increasing compliance with well‐child visits, taking advantage of nonwell‐child visits, and implementing Medicaid policies that allow for optimal timing of visits.
Dental Fluorosis in 12‐ and 15‐Year‐Olds at High Altitudes in Above‐Optimal Fluoridated Communities in Mexico
Authors: Pontigo‐Loyola América P. , Islas‐Márquez Arturo , Loyola‐Rodríguez Juan P. , Maupome Gerardo , Marquez‐Corona M. Lourdes , Medina‐Solis Carlo E.
Published: September 2008

Objective: The purpose of this study was to determine the prevalence and severity of dental fluorosis in Mexican adolescents. Materials and Methods: A cross‐sectional epidemiological study was carried out in 1,024 adolescents 12 and 15 years old residing in three naturally fluoridated locales at high altitudes above sea level (>2,000 m or >6,560 ft) in Tula de Allende, Hidalgo, Mexico. Participants had lived in those communities from birth to their sixth birthday. Both the Modified Dean Index and the Community Fluorosis Index were calculated. Results: The overall fluorosis prevalence was 83.8 percent. Fluorosis prevalence in El Llano (3.07 ppmF), San Marcos (1.38 ppmF), and Tula Centro (1.42 ppmF) was 94.7, 89.8, and 81.9 percent, respectively. Overall, the Community Fluorosis Index was 1.85. We observed a high prevalence of dental fluorosis – mostly very mild (35.9 percent), but also uncommonly severe (20.6 percent). Conclusions: At least 8 out of 10 adolescents had some level of fluorosis, and such prevalence is considered to be a dental public health problem. A relationship between fluoride concentration in water in each community and fluorosis was observed. The high fluorosis prevalence and severity might possibly be associated with the high altitude of the communities.
A Phase 3, Randomized, Placebo‐controlled Trial of Filgrastim in Patients with Haematological Malignancies Undergoing Matched‐related Allogeneic Bone Marrow Transplantation
Authors: Ernst Peter , Bacigalupo Andrea , Ringdén Olle , Ruutu Tapani , Kolb Hans J. , Lawrinson Susan , Skacel Tomas
Published: September 2008

Introduction.  Recombinant granulocyte colony‐stimulating factor (G‐CSF) may aid engraftment post high‐dose chemo‐/radiotherapy in patients with haematological malignancies undergoing allogeneic bone marrow transplantation (BMT); however, the effects of G‐CSF on graft‐versus‐host disease (GvHD), relapse, and survival are not well defined.

Methods.  In this double‐blind, randomized, placebo‐controlled, multicentre, phase 3 study, the effects of the G‐CSF Filgrastim on neutrophil and platelet recovery, and on clinical outcomes were evaluated. Patients (12–55 years) receiving an allogeneic BMT for a haematological malignancy were randomized to receive Filgrastim 5 µg/kg or placebo. Study treatment was continued until patients achieved an absolute neutrophil count (ANC) ≥0.5 × 109/L, or until day 42.

Results.  Fifty‐one patients (Filgrastim, N = 25; placebo, N = 26) were evaluable. Patients treated with Filgrastim had significantly faster engraftment with ANC ≥0.5 × 109/L being achieved after a median (range) of 15.0 (1.0–22.0) days vs. 19.0 (15.0–28.0) days for placebo (P < 0.0001). The incidence of GvHD was comparable for both groups. During the limited follow‐up (2 years), Filgrastim had no adverse effect on mortality and possibly reduced the rate of relapse.
Microsatellite markers for eastern hemlock (Tsuga canadensis)
Authors: SHAMBLIN BRIAN M. , FAIRCLOTH BRANT C. , JOSSERAND SEDLEY A. , NELSON C. DANA , NAIRN CAMPBELL J.
Published: September 2008

We describe polymerase chain reaction primer pairs and reaction conditions for amplification of 15 microsatellite loci from eastern hemlock (Tsuga canadensis). The primers were tested on 23 individuals from a natural population in southwestern North Carolina, USA. These primers yielded an average of 5.9 alleles per locus (range of 2–14), an average observed heterozygosity of 0.45 (range 0.14–0.73), and an average polymorphic information content of 0.54 (range 0.28–0.86). In addition, eight of the primer pairs were found to amplify microsatellite loci in one or more additional species of Tsuga.
Development of microsatellite markers inAnnona crassifloraMart., a Brazilian Cerrado fruit tree species
Authors: PEREIRA M. F. , BANDEIRA L. F. , BLANCO A. J. V. , CIAMPI A. Y. , COELHO A. S. G.
Published: September 2008

Annona crassifloraMart. (Annonaceae) is a native fruit species of the region of Brazilian Cerrado with a high agronomic potential, although without any traces of domestication. A set of 10 microsatellite primer pairs was developed from an enriched genome library (TC13). An average of 19.3 alleles per locus was detected. Observed heterozygosity estimates were consistently lower than those obtained for gene diversity, evidencing a departure from Hardy–Weinberg expected proportions. The reported set of markers showed to be highly informative and constitutes a powerful tool for the development of genetic characterization studies inA. crassiflora.
Isolation and characterization of microsatellite markers for Carolina hemlock (Tsuga caroliniana)
Authors: JOSSERAND S. A. , POTTER K. M. , ECHT C. S. , NELSON C. D.
Published: September 2008

We describe the isolation and characterization of 31 polymorphic di‐ and trinucleotide microsatellite marker loci for Carolina hemlock (Tsuga caroliniana Englem.). In addition, primer pairs for 16 loci amplified scoreable alleles in six other Tsuga species. In eastern North America, both Carolina hemlock and eastern hemlock (Tsuga canadensis [L.] Carr.) populations are declining due to infestation by hemlock woolly adelgid, Adelges tsugae. The markers described here should enhance population genetic studies of hemlocks, providing valuable information for conserving and restoring these important forest tree species.
Twenty‐three microsatellite DNA loci for population genetic studies and parentage assignment in orangethroat darter, Etheostoma spectabile
Authors: HUDMAN S. P. , GROSE M. J. , LANDIS J. B. , SKALSKI G. T. , WILEY E. O.
Published: September 2008

The genus Etheostoma is a species‐rich and ecologically important group of fishes in North America. The orangethroat darter (Etheostoma spectabile) is widely distributed and abundant in headwater streams throughout the central Midwest, and is an excellent model for ecological and mating system studies. We developed 23 novel, polymorphic, and independent microsatellite loci for E. spectabile. We found from two to 14 alleles per locus, and observed heterozygosities ranged from 0.39 to 1.0. These markers, in combination with others isolated from Etheostoma taxa, will be useful for ecological and evolutionary studies in the genus.
Characterization of new microsatellite markers derived from sequence databases for the emu (Dromaius novaehollandiae)
Authors: YÁÑEZ JOSÉ M. , GONZÁLEZ RUTH , ANGULO JENNIFFER , VIDAL RODRIGO , SANTOS JOSÉ L. , MARTÍNEZ VICTOR
Published: September 2008

The emu (Dromaius novaehollandiae), a member of ratite family, is native to Australia and has been introduced to other countries worldwide. In this work, 10 polymorphic microsatellite loci were isolated and characterized for emu from public sequences. Polymorphism was surveyed in 22 individuals from two different populations kept in captivity. Between two and 11 alleles were found per locus, and the observed heterozygosity ranged from 0.05 to 0.85, in accordance with expectations. These markers will be useful as tools for detecting levels of genetic variation, reconstructing pedigrees (for quantitative genetic analysis) and identifying markers associated to fitness traits in emu populations.
Increased anterior chamber penetration of topical levofloxacin 0.5% after pulsed dosing in cataract patients
Authors: Sundelin Karin , Seal David , Gardner Susanne , Ropo Auli , Geldsetzer Klaus , Lokkila Jukka , Stenevi Ulf
Published: September 2008

Purpose:  This study aimed to investigate the rise in aqueous humour (AH) levels of levofloxacin after a specific perioperative pulsed topical drop regimen.

Methods:  Thirty patients undergoing phacoemulsification surgery were administered two preoperative drops of levofloxacin 0.5%, 30 mins apart, and three pulsed drops postoperatively, 5 mins apart. Aqueous humour levels of levofloxacin were measured at the start of surgery and from 5 mins to 90 mins after the last postoperative drop. Samples from individual patients were collected at the time of surgery and at one additional sampling interval by aqueous tap, and analysed using a high‐performance liquid chromatography assay.

Results:  Aqueous humour levels of levofloxacin continued to rise gradually, reaching a mean peak level (Cmax) of 4.4 μg/ml (± 2.5) at 60 mins after the last postoperative drop was administered. This level exceeded the minimum inhibitory concentration of common ocular pathogens at least fourfold. At 90 mins after the last drop, mean AH levels remained > 3 μg/ml.

Conclusions:  This is the first study to measure AH levels of levofloxacin after postoperative pulsed dosing in humans. Higher AH levels were found than in previously reported studies in which only preoperative drops were given and levels were measured at the time of surgery. Levels of levofloxacin continued to rise for 60 mins after administration of the last postoperative drop, demonstrating that delivery and maintenance of effective antibiotic levels may be achievable with alternative dosing schedules.
“True” as Ambiguous
Authors: KÖLBEL MAX
Published: September 2008

In this paper, I argue (a) that the predicate “true” is ambiguously used to express a deflationary and a substantial concept of truth and (b) that the two concepts are systematically related in that substantial truths are deflationary truths of a certain kind. Claim (a) allows one to accept the main insights of deflationism but still take seriously, and participate in, the traditional debate about the nature of truth. Claim (b) is a contribution to that debate. The overall position is not new and it has previously been defended by supervaluationists about vagueness. However, the position is here motivated in a new, independent way, and an explanation is offered why some uses of “true” do not seem to require disambiguation.
Chances, Counterfactuals, and Similarity
Authors: WILLIAMS J. ROBERT G.
Published: September 2008

John Hawthorne in a recent paper takes issue with Lewisian accounts of counterfactuals, when relevant laws of nature are chancy. I respond to his arguments on behalf of the Lewisian, and conclude that while some can be rebutted, the case against the original Lewisian account is strong.

I develop a neo‐Lewisian account of what makes for closeness of worlds. I argue that my revised version avoids Hawthorne's challenges. I argue that this is closer to the spirit of Lewis's first (non‐chancy) proposal than is Lewis's own suggested modification.
Respect for Just Revenge
Authors: ROSEBURY BRIAN
Published: September 2008

The paper considers acts of private (in the sense of individually motivated and extra‐legal) revenge, and draws attention to a special kind of judgement we may make of such acts. While endorsing the general view that an act of private revenge must be morally wrong, it maintains that under certain special conditions (which include its being just) it is susceptible of a rational respect from others which is based on its standing outside morality, as a choice by the revenger not to act morally but to obey other compelling motives. This thesis is tested against various objections, notably those which doubt the intelligibility or application of such non‐moral ‘respect,’ or would assimilate it to moral approval; and it is distinguished from various positions with which it might be confused, such as the ‘admirable immorality’ of Slote, or the Nietzschean critique of morality.
The Epistemological Role of Episodic Recollection
Authors: SOTERIOU MATTHEW
Published: September 2008

In what respects is episodic recollection active, and subject to the will, like perceptual imagination, and in what respects is it passive, like perception, and how do these matters relate to its epistemological role? I present an account of the ontology of episodic recollection that provides answers to these questions. According the account I recommend, an act of episodic recollection is not subject to epistemic evaluation—it is neither justified nor unjustified—but it can provide one with a distinctive source of warrant for judgements about the past when it is accompanied by knowledge that one is recollecting, as well as knowledge of what one is recollecting. While the account concedes that when one recollects one’s attitude to what is recollected cannot be one of observation, it nevertheless accommodates the notion that episodic recollection involves a form of mental time‐travel—a case of re‐visiting, or re‐acquaintance with, some past episode.
Measurement of interaction forces between fibrinogen coated probes and mica surface with the atomic force microscope: The pH and ionic strength effect
Authors: Tsapikouni Theodora S. , Allen Stephanie , Missirlis Yannis F.
Published: September 2008

The study of protein-surface interactions is of great significance in the design of biomaterials and the evaluation of molecular processes in tissue engineering. The authors have used atomic force microscopy (AFM) to directly measure the force of attraction/adhesion of fibrinogen coated tips to mica surfaces and reveal the effect of the surrounding solution pH and ionic strength on this interaction. Silica colloid spheres were attached to the AFM cantilevers and, after plasma deposition of poly(acrylic acid), fibrinogen molecules were covalently bound on them with the help of the cross-linker 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) in the presence of N-hydroxysulfosuccinimide (sulfo-NHS(. The measurements suggest that fibrinogen adsorption is controlled by the screening of electrostatic repulsion as the salt concentration increases from 15 to 150 mM, whereas at higher ionic strength (500 mM) the hydration forces and the compact molecular conformation become crucial, restricting adsorption. The protein attraction to the surface increases at the isoelectric point of fibrinogen (pH 5.8), compared with the physiological pH. At pH 3.5, apart from fibrinogen attraction to the surface, evidence of fibrinogen conformational changes is observed, as the pH and the ionic strength are set back and forth, and these changes may account for fibrinogen aggregation in the protein solution at this pH.
Calculation of adsorption free energy for solute-surface interactions using biased replica-exchange molecular dynamics
Authors: Wang Feng , Stuart Steven J. , Latour Robert A.
Published: September 2008

The adsorption behavior of a biomolecule, such as a peptide or protein, to a functionalized surface is of fundamental importance for a broad range of applications in biotechnology. The adsorption free energy for these types of interactions can be determined from a molecular dynamics simulation using the partitioning between adsorbed and nonadsorbed states, provided that sufficient sampling of both states is obtained. However, if interactions between the solute and the surface are strong, the solute will tend to be trapped near the surface during the simulation, thus preventing the adsorption free energy from being calculated by this method. This situation occurs even when using an advanced sampling algorithm such as replica-exchange molecular dynamics (REMD). In this paper, the authors demonstrate the fundamental basis of this problem using a model system consisting of one sodium ion (Na+) as the solute positioned over a surface functionalized with one negatively charged group (COO-) in explicit water. With this simple system, the authors show that sufficient sampling in the coordinate normal to the surface cannot be obtained by conventional REMD alone. The authors then present a method to overcome this problem through the use of an adaptive windowed-umbrella sampling technique to develop a biased-energy function that is combined with REMD. This approach provides an effective method for the calculation of adsorption free energy for solute-surface interactions.
Advanced nanostructures for the control of biofouling: The FP6 EU Integrated Project AMBIO
Authors: Rosenhahn Axel , Ederth Thomas , Pettitt Michala E.
Published: September 2008

The colonization of man made structures by marine or freshwater organisms or “biofouling” is a problem for maritime and aquaculture industries. Increasing restrictions on the use of toxic coatings that prevent biofouling, create a gap in the market that requires new approaches to produce novel nonbiocidal alternatives. This review details the systematic strategy adopted by an FP6 EU Integrated Project “AMBIO” to develop fundamental understanding of key surface properties that influence settlement and adhesion of fouling organisms. By this approach the project contributes to the understanding of fundamental phenomena involved in biofouling, and to the development of environmentally benign solutions by coating manufacturers within the consortium. © 2008 American Vacuum Society.
Effect of Therapeutic Chemical Agents In Vitro and on Experimental Meningoencephalitis Due toNaegleria fowleri
Authors: Kim Jong-Hyun , Jung Suk-Yul , Lee Yang-Jin , Song Kyoung-Ju , Kwon Daeho , Kim Kyongmin , Park Sun , Im Kyung-Il , Shin Ho-Joon
Published: September 2008

Naegleria fowleriis a ubiquitous, pathogenic free-living amoeba; it is the most virulentNaegleriaspecies and causes primary amoebic meningoencephalitis (PAME) in laboratory animals and humans. Although amphotericin B is currently the only agent available for the treatment of PAME, it is a very toxic antibiotic and may cause many adverse effects on other organs. In order to find other potentially therapeutic agents forN. fowleriinfection, the present study was undertaken to evaluate the in vitro and in vivo efficacies of miltefosine and chlorpromazine against pathogenicN. fowleri. The result showed that the growth of the amoeba was effectively inhibited by treatment with amphotericin B, miltefosine, and chlorpromazine. WhenN. fowleritrophozoites were treated with amphotericin B, miltefosine, and chlorpromazine, the MICs of the drug were 0.78, 25, and 12.5 μg/ml, respectively, on day 2. In experimental meningoencephalitis of mice that is caused byN. fowleri, the survival rates of mice treated with amphotericin B, miltefosine, and chlorpromazine were 40, 55, and 75%, respectively, during 1 month. The average mean time to death for the amphotericin B, miltefosine, and chlorpromazine treatments was 17.9 days. In this study, the effect of drugs was found to be optimal when 20 mg/kg was administered three times on days 3, 7, and 11. Finally, chlorpromazine had the best therapeutic activity againstN. fowleriin vitro and in vivo. Therefore, it may be a more useful therapeutic agent for the treatment of PAME than amphotericin B.
Use of Gyrase Resistance Mutants To Guide Selection of 8-Methoxy-Quinazoline-2,4-Diones
Authors: German Nadezhda , Malik Muhammad , Rosen Jonathan D. , Drlica Karl , Kerns Robert J.
Published: September 2008

A series of 1-cyclopropyl-8-methoxy-quinazoline-2,4-diones was synthesized and evaluated for lowering the ratio of the antimicrobial MIC in gyrase resistance mutants to that in the
gyr
+
(wild type) using isogenic strains of
Escherichia coli
. Dione features that lowered this ratio were a 3-amino group and C-7 ring structure (3-aminomethyl pyrrolidinyl < 3-aminopyrrolidinyl < diazobicyclo < 2-ethyl piperazinyl). The wild-type MIC was also lowered. With the most active derivative tested, many
gyrA
resistance mutant types were as susceptible as, or more susceptible than, wild-type cells. The most active 2,4-dione derivatives were also more active with two quinolone-resistant
gyrB
mutants than with wild-type cells. With respect to lethality, the most bacteriostatic 2,4-dione killed
E. coli
at a rate that was affected little by a
gyrA
resistance mutation, and it exhibited a rate of killing similar to its cognate fluoroquinolone at 10× the MIC. Population analysis with wild-type
E. coli
applied to agar showed that the mutant selection window for the most active 2,4-dione was narrower than that for the cognate fluoroquinolone or for ciprofloxacin. These data illustrate a new approach to guide early-stage antimicrobial selection. Use of antimutant activity (i.e., ratio of the antimicrobial MIC in a mutant strain to the antimicrobial MIC in a wild-type strain) as a structure-function selection criterion can be combined with traditional efforts aimed at lowering antimicrobial MICs against wild-type organisms to more effectively afford lead molecules with activity against both wild-type and mutant cells.
Interspecies Recombination Occurs Frequently in Quinolone Resistance-Determining Regions of Clinical Isolates of
Streptococcus pyogenes
Authors: Duesberg Christoph B. , Malhotra-Kumar Surbhi , Goossens Herman , McGee Lesley , Klugman Keith P. , Welte Tobias , Pletz Mathias W. R.
Published: September 2008

Fluoroquinolone resistance in
Streptococcus pyogenes
has been reported only anecdotally, but a recent Belgian surveillance study found a rate of nonsusceptibility of 5.4%. From an analysis of these isolates, we show that interspecies horizontal gene transfer within the
parC
quinolone resistance-determining region is a frequent phenomenon that might contribute to fluoroquinolone resistance.
Repeated Administration of High-Dose Intermittent Rifapentine Reduces Rifapentine and Moxifloxacin Plasma Concentrations
Authors: Dooley Kelly , Flexner Charles , Hackman Judith , Peloquin Charles A. , Nuermberger Eric , Chaisson Richard E. , Dorman Susan E.
Published: September 2008

Moxifloxacin- and rifapentine-based regimens are under investigation for the treatment of tuberculosis. However, rifapentine may induce enzymes that metabolize moxifloxacin, resulting in decreased moxifloxacin concentrations. In this phase I, two-period, sequential-design study, 13 subjects received 400 mg moxifloxacin daily for 4 days followed by daily moxifloxacin coadministered with 900 mg rifapentine thrice weekly. Pharmacokinetic analyses were performed after the 4th and 19th doses of moxifloxacin and after the 1st and 7th doses of rifapentine. For moxifloxacin, the mean area under the concentration-time curve from 0 to 24 h (AUC
0-24
) decreased by 17.2% (
P
= 0.0006) when the drug was coadministered with rifapentine, and the mean half-life (
t
1/2
) decreased from 11.1 to 8.9 h (
P
= 0.0033). For rifapentine, the mean AUC
0-48
after seven thrice-weekly doses decreased by 20.3% (
P
= 0.0035) compared to the AUC
0-48
after the first dose, and the mean
t
1/2
decreased from 18.5 to 14.8 h (
P
= 0.0004). The AUC
0-48
for the 25-desacetyl-rifapentine metabolite diminished 21%. Two days after completing the study drugs, one subject developed a fever and hepatitis, and another developed a flu-like illness with a rash. In conclusion, rifapentine modestly reduced moxifloxacin concentrations. Changes consistent with rifapentine autoinduction of metabolism were seen. Adverse reactions in two subjects may have represented rifamycin hypersensitivity syndrome, although some features were atypical.
Hitchhiking and Selective Sweeps of
Plasmodium falciparum
Sulfadoxine and Pyrimethamine Resistance Alleles in a Population from Central Africa
Authors: McCollum Andrea M. , Basco Leonardo K. , Tahar Rachida , Udhayakumar Venkatachalam , Escalante Ananias A.
Published: September 2008

Sulfadoxine-pyrimethamine (SP) resistance in
Plasmodium falciparum
is encoded by a number of mutations in the dihydrofolate reductase (
dhfr
) and dihydropteroate synthetase (
dhps
) genes. Here, we have characterized point mutations in
dhfr
and
dhps
and microsatellite loci around
dhfr
on chromosome 4 and
dhps
on chromosome 8 as well as neutral markers on chromosomes 2 and 3 in 332 samples from Yaoundé, Cameroon. The triple mutant
dhfr
haplotype that originated in Southeast Asia is the most predominant in this sample set, but we also find additional independent haplotypes at low frequency and an incipient process of genetic differentiation among alleles of Southeast Asian origin. As reported for other African populations, we find evidence of a selective sweep for resistant
dhfr
mutants in this Cameroonian population due to drug selection. Although we find evidence for a selective sweep in
dhps
mutants associated with SP resistance, the dynamics of
dhps
mutants appear different than those observed for
dhfr
mutants. Overall, our results yield support for the use of microsatellite markers to track resistant parasites; however, the detection of resistant
dhfr
alleles in low frequency, the evidence of divergence among
dhfr
alleles that share a common evolutionary origin, and the distinct dynamics of resistant
dhps
alleles emphasize the importance of comprehensive, population-based investigations to evaluate the effects of drug selection on parasite populations.
Assessing the Efficacy of Cidofovir against Herpesvirus-Induced Genital Lesions in Goats Using Different Therapeutic Regimens
Authors: Tempesta M. , Crescenzo G. , Camero M. , Bellacicco A. L. , Tarsitano E. , Decaro N. , Neyts J. , Martella V. , Buonavoglia C.
Published: September 2008

Caprine herpesvirus 1 (CpHV-1) infection in goats induces genital vesicular-ulcerative lesions that strictly resemble those produced by human herpesvirus 2 in humans. In previous studies, the potent inhibition of CpHV-1 by cidofovir was demonstrated. Cidofovir antiherpetic activity was evaluated in goats infected experimentally by the vaginal route with CpHV-1 and then treated locally at different times after infection. The administration of 1% cidofovir cream onto vaginal mucosa was able to prevent the onset of genital lesions and to decrease significantly the titers of the virus shed by the infected animals, notably in the groups treated shortly after infection (24 and 48 h). The efficacy of cidofovir against caprine herpesvirus infection was higher when the treatment was started shortly after infection than when lesions were already present and advanced. Herpesvirus genital infection of goats is a useful animal model to study the activity of antiviral drugs against human herpesvirus infections.
KHM-1, a Novel Plasmid-Mediated Metallo-β-Lactamase from a
Citrobacter freundii
Clinical Isolate
Authors: Sekiguchi Jun-ichiro , Morita Koji , Kitao Tomoe , Watanabe Noboru , Okazaki Mitsuhiro , Miyoshi-Akiyama Tohru , Kanamori Masato , Kirikae Teruo
Published: September 2008

A novel gene,
bla
KHM-1
, encoding a metallo-β-lactamase, KHM-1, was cloned from a clinical isolate of
Citrobacter freundii
resistant to most β-lactam antibiotics.
Escherichia coli
expressing
bla
KHM-1
was resistant to all broad-spectrum β-lactams except for monobactams and showed reduced susceptibility to carbapenems. Recombinant KHM-1 exhibited EDTA-inhibitable hydrolytic activity against most β-lactams, with an overall preference for cephalosporins.
In Vitro Activities of ER-119884 and E5700, Two Potent Squalene Synthase Inhibitors, against
Leishmania amazonensis
: Antiproliferative, Biochemical, and Ultrastructural Effects
Authors: Fernandes Rodrigues Juliany Cola , Concepcion Juan Luis , Rodrigues Carlos , Caldera Aura , Urbina Julio A. , de Souza Wanderley
Published: September 2008

ER-119884 and E5700, novel arylquinuclidine derivatives developed as cholesterol-lowering agents, were potent in vitro growth inhibitors of both proliferative stages of
Leishmania amazonensis
, the main causative agent of cutaneous leishmaniasis in South America, with the 50% inhibitory concentrations (IC
50
s) being in the low-nanomolar to subnanomolar range. The compounds were very potent noncompetitive inhibitors of native
L. amazonensis
squalene synthase (SQS), with inhibition constants also being in the nanomolar to subnanomolar range. Growth inhibition was strictly associated with the depletion of the parasite's main endogenous sterols and the concomitant accumulation of exogenous cholesterol. Using electron microscopy, we identified the intracellular structures affected by the compounds. A large number of lipid inclusions displaying different shapes and electron densities were observed after treatment with both SQS inhibitors, and these inclusions were associated with an intense disorganization of the membrane that surrounds the cell body and flagellum, as well as the endoplasmic reticulum and the Golgi complex. Cells treated with ER-119884 but not those treated with E5700 had an altered cytoskeleton organization due to an abnormal distribution of tubulin, and many were arrested at cytokinesis. A prominent contractile vacuole and a phenotype typical of programmed cell death were frequently found in drug-treated cells. The selectivity of the drugs was demonstrated with the JC-1 mitochondrial fluorescent label and by trypan blue exclusion tests with macrophages, which showed that the IC
50
s against the host cells were 4 to 5 orders of magnitude greater that those against the intracellular parasites. Taken together, our results show that ER-119884 and E5700 are unusually potent and selective inhibitors of the growth of
Leishmania amazonensis
, probably because of their inhibitory effects on de novo sterol biosynthesis at the level of SQS, but some of our observations indicate that ER-119884 may also interfere with other cellular processes.
Vaccination of Mice with a Yop Translocon Complex Elicits Antibodies That Are Protective against Infection with F1
−
Yersinia pestis
Authors: Ivanov Maya I. , Noel Betty L. , Rampersaud Ryan , Mena Patricio , Benach Jorge L. , Bliska James B.
Published: September 2008

Yersinia pestis
, the bacterial agent of plague, secretes several proteins important for pathogenesis or host protection. The F1 protein forms a capsule on the bacterial cell surface and is a well-characterized protective antigen but is not essential for virulence. A type III secretion system that is essential for virulence exports Yop proteins, which function as antiphagocytic or anti-inflammatory factors. Yop effectors (e.g., YopE) are delivered across the host cell plasma membrane by a translocon, composed of YopB and YopD. Complexes of YopB, YopD, and YopE (BDE) secreted by
Yersinia pseudotuberculosis
were purified by affinity chromatography and used as immunogens to determine if antibodies to the translocon could provide protection against
Y. pestis
in mice. Mice vaccinated with BDE generated high-titer immunoglobulin G antibodies specific for BDE, as shown by enzyme-linked immunosorbent assay and immunoblotting, and were protected against lethal intravenous challenge with F1
−
but not F1
+
Y. pestis.
Mice passively immunized with anti-BDE serum were protected from lethal challenge with F1
−
Y. pestis
. The YopB protein or a complex of YopB and YopD (BD) was purified and determined by vaccination to be immunogenic in mice. Mice actively vaccinated with BD or passively vaccinated with anti-BD serum were protected against lethal challenge with F1
−
Y. pestis
. These results indicate that anti-translocon antibodies can be used as immunotherapy to treat infections by F1
−
Y. pestis
.
Dynamin-2-Dependent Targeting ofMannheimia haemolyticaLeukotoxin to Mitochondrial Cyclophilin D in Bovine Lymphoblastoid Cells
Authors: Atapattu Dhammika N. , Albrecht Ralph M. , McClenahan David J. , Czuprynski Charles J.
Published: September 2008

Exotoxins which belong to the family containing the RTX toxins (repeats in toxin) contribute to a variety of important human and animal diseases. One example of such a toxin is the potent leukotoxin (LKT) produced by the bovine respiratory pathogenMannheimia haemolytica.LKT binds to CD18, resulting in the death of bovine leukocytes. In this study, we showed that internalized LKT binds to the outer mitochondrial membrane, which results in the release of cytochromecand collapse of the mitochondrial membrane potential (ψm). Incubation of bovine lymphoblastoid cells (BL-3 cells) with the mitochondrial membrane-stabilizing agent cyclosporine (CSA) reduced LKT-mediated cytotoxicity, cytochromecrelease, and collapse of the ψm. Coimmunoprecipitation and intracellular binding studies suggested that LKT binds to the mitochondrial matrix protein cyclophilin D. We also demonstrated that LKT mobilizes the vesicle scission protein dynamin-2 from mitochondria to the cell membrane. Incubation with CSA depleted mitochondrial dynamin-2 in BL-3 cells, making it unavailable for vesicle scission and LKT internalization. The results of this study show that LKT trafficking and LKT-mediated cell death involve dynamin-2 and cyclophilin D, in a process that can be prevented by the mitochondrial membrane-protecting function of CSA.
Functional Mapping of YadA- and Ail-Mediated Binding of Human Factor H toYersinia enterocoliticaSerotype O:3
Authors: Biedzka-Sarek Marta , Salmenlinna Saara , Gruber Markus , Lupas Andrei N. , Meri Seppo , Skurnik Mikael
Published: September 2008

Yersinia enterocoliticais an enteric pathogen that exploits diverse means to survive in the human host. UponY. enterocoliticaentry into the human host, bacteria sense and respond to variety of signals, one of which is the temperature. Temperature in particular has a profound impact onY. enterocoliticagene expression, as most of its virulence factors are expressed exclusively at 37°C. These include two outer membrane proteins, YadA and Ail, that function as adhesins and complement resistance (CR) factors. Both YadA and Ail bind the functionally active complement alternative pathway regulator factor H (FH). In this study, we characterized regions on both proteins involved in CR and the interaction with FH. Twenty-eight mutants having short (7 to 41 amino acids) internal deletions within the neck and stalk of YadA and two complement-sensitive site-directed Ail mutants were constructed to map the CR and FH binding regions of YadA and Ail. Functional analysis of the YadA mutants revealed that the stalk of YadA is required for both CR and FH binding and that FH appears to target several conformational and discontinuous sites of the YadA stalk. On the other hand, the complement-sensitive Ail mutants were not affected in FH binding. Our results also suggested that Ail- and YadA-mediated CR does not depend solely on FH binding.
Bacterial Conjugation in the Cytoplasm of Mouse Cells
Authors: Lim Yin Mei , de Groof Ad J. C. , Bhattacharjee Mrinal K. , Figurski David H. , Schon Eric A.
Published: September 2008

Intracellular pathogenic organisms such as salmonellae and shigellae are able to evade the effects of many antibiotics because the drugs are not able to penetrate the plasma membrane. In addition, these bacteria may be able to transfer genes within cells while protected from the action of drugs. The primary mode by which virulence and antibiotic resistance genes are spread is bacterial conjugation. Salmonellae have been shown to be competent for conjugation in the vacuoles of cultured mammalian cells. We now show that the conjugation machinery is also functional in the mammalian cytosol. Specially constructed
Escherichia coli
strains expressing
Shigella flexneri
plasmid and chromosomal virulence factors for escape from vacuoles and synthesizing the invasin protein from
Yersinia pseudotuberculosis
to enhance cellular entry were able to enter 3T3 cells and escape from the phagocytic vacuole. One bacterial strain (the donor) of each pair to be introduced sequentially into mammalian cells had a conjugative plasmid. We found that this plasmid could be transferred at high frequency. Conjugation in the cytoplasm of cells may well be a general phenomenon.
Epithelial and Mesenchymal Cells in the Bovine Colonic Mucosa Differ in Their Responsiveness toEscherichia coliShiga Toxin 1
Authors: Stamm Ivonne , Mohr Melanie , Bridger Philip S. , Schröpfer Elmar , König Matthias , Stoffregen William C. , Dean-Nystrom Evelyn A. , Baljer Georg , Menge Christian
Published: September 2008

Bovine colonic crypt cells express CD77 molecules that potentially act as receptors for Shiga toxins (Stx). The implication of this finding for the intestinal colonization of cattle by human pathogenic Stx-producingEscherichia coli(STEC) remains undefined. We used flow cytometric and real-time PCR analyses of primary cultures of colonic crypt cells to evaluate cell viability, CD77 expression, and gene transcription in the presence and absence of purified Stx1. A subset of cultured epithelial cells had Stx receptors which were located mainly intracellularly, with a perinuclear distribution, and were resistant to Stx1-induced apoptosis and Stx1 effects on chemokine expression patterns. In contrast, a population of vimentin-positive cells, i.e., mesenchymal/nonepithelial cells that had high numbers of Stx receptors on their surface, was depleted from the cultures by Stx1. In situ, CD77+cells were located in the lamina propria of the bovine colon by using immunofluorescence staining. A newly established vimentin-positive crypt cell line with high CD77 expression resisted the cytolethal effect of Stx1 but responded to Stx1 with a significant increase in interleukin-8 (IL-8), GRO-α, MCP-1, and RANTES mRNA. Combined stimulation with lipopolysaccharide and Stx1 increased IL-10 mRNA. Our results show that bovine colonic crypt cells of epithelial origin are resistant to both the cytotoxic and modulatory effects of Stx1. In contrast, some mucosal mesenchymal cells, preliminarily characterized as mucosal macrophages, are Stx1-responsive cells that may participate in the interaction of STEC with the bovine intestinal mucosa.