bluuebunny/crossref_metadata_2025 huggingface dataset:
Take‐home naloxone to reduce heroin death
Authors: Baca Catherine T. , Grant Kenneth J.
Published: November 2005

Background This paper reviews the relevant literature related to the distribution of take‐home naloxone.

Methods A Medline search was conducted on articles published between January 1990 and June 2004 to identify scientific literature relevant to this subject. Those publications were reviewed, and from them other literature was identified and reviewed.

Results The prevalence, pathophysiology and circumstances of heroin overdose, and also bystander response are included in this review. Naloxone peer distribution has been instituted to varying degrees in the United States, Italy, Spain, Germany and the United Kingdom.

Conclusion At this point the evidence supporting naloxone distribution is primarily anecdotal, although promising. Although the distribution of naloxone holds promise for further reducing heroin overdose mortality, problems remain. Naloxone alone may be insufficient in some cases to revive the victim, and cardiopulmonary resuscitation (CPR), especially rescue breathing, may also be needed. A second dose of naloxone might be necessary. Complications following resuscitation from overdose may infrequently need in‐hospital care. Mortality from injecting without anyone else present will be unaffected by take‐home naloxone. Take‐home naloxone should be studied in a rigorous scientific manner.
Phytophthora cinnamomi
Authors: HARDHAM ADRIENNE R.
Published: November 2005

Phytophthora cinnamomi Rands was first isolated from cinnamon trees in Sumatra in 1922. The pathogen is believed to have originated near Papua New Guinea but now has a worldwide distribution. P. cinnamomi is heterothallic with A1 and A2 mating types; however, even in areas in which both mating types are present, it appears that genetic diversity arises asexually rather than as a result of sexual recombination. P. cinnamomi can grow saprophytically in the soil for long periods, rapidly capitalizing on the advent of favourable conditions to sporulate and produce vast numbers of asexual, biflagellate zoospores. The motile zoospores are attracted to suitable infection sites, where they attach and invade the plant. Within a few days, hyphae ramify throughout the tissues of susceptible plants, forming sporangia on the plant surface and rapidly amplifying the disease inoculum. Over the last 10–15 years, molecular analyses have clarified details of the phylogeny of P. cinnamomi and other Oomycetes. Research on P. cinnamomi has given rise to a more comprehensive understanding of the structure and function of the motile zoospores. New methods have been developed for P. cinnamomi identification and diagnosis. Long‐term studies of diseased sites, particular those in southern Australia, have produced a better understanding of the epidemiology of P. cinnamomi diseases. Research has also increased our knowledge of the mode of action and efficacy of inhibitors of P. cinnamomi diseases, especially the phosphonates. This review will present an overview of the advances these studies have made in our understanding of P. cinnamomi pathogenicity, epidemiology and control.

 Taxonomy: Phytophthora cinnamomi Rands; kingdom Chromista; phylum Oomycota; order Peronosporales; family Peronosporaceae; genus Phytophthora.

 Host range: Likely to infect in excess of 3000 species of plants including over 2500 Australian native species, and crops such as avocado, pineapple, peach, chestnut and macadamia.

 Disease symptoms: A root pathogen which usually causes rotting of fine and fibrous roots but which can also cause stem cankers. Often causes dieback of young shoots and is thought to do so as a result of interference with transpiration from roots to shoots.

 Useful websites: http://genome.jgi‐psf.org/physo00.info.html; http://phytophthora.vbi.vt.edu.
Review article: similarities and differences among delayed‐release proton‐pump inhibitor formulations
Authors: HORN J. R. , HOWDEN C. W.
Published: November 2005

Proton‐pump inhibitors are acid‐labile, and require an enteric coating to protect them from degradation in the stomach when given orally. However, this leads to delayed absorption and onset of action of the proton‐pump inhibitor.

This article aims to review the similarities and differences between the various formulations of delayed release proton‐pump inhibitors. Delayed‐release omeprazole and delayed‐release lansoprazole have been suspended in sodium bicarbonate for tube administration; however, for omeprazole, absorption is further impaired and antisecretory effects are disappointing. Although such formulations may be more convenient for clinical use in certain patient groups, absorption of the proton‐pump inhibitor is still influenced by residual enteric coating. There are few differences among the currently available delayed‐release proton‐pump inhibitors with respect to their pharmacodynamic effects during chronic administration. There are minor formulation‐based pharmacokinetic differences among these agents, primarily reflected in their bioavailability following the first few doses.

Differences in bioavailability may explain slight differences in the rate of onset of maximal antisecretory effect. However, minor pharmacodynamic and pharmacokinetic differences are not associated with meaningful differences in clinical outcomes.
Raptorial birds and environmental gradients in the southern Neotropics: A test of species‐richness hypotheses
Authors: BELLOCQ M. I. , GÓMEZ‐INSAUSTI R.
Published: November 2005

Abstract  We investigated the spatial patterns of raptor species richness in the southern Neotropics and tested three hypotheses that were most likely to explain spatial variations: ambient energy, productivity and habitat heterogeneity. We used non‐linear regression analysis and eliminated alternative hypotheses by finding the best single environmental predictor of raptor species richness among potential evapotranspiration (PET), actual evapotranspiration (AET), mean annual temperature and precipitation and vegetation structure coefficient. As expected, the number of raptor species decreases monotonically as latitude increases. Raptor species richness was significantly correlated with each of the environmental factors considered in this study, reflecting covariation of climatic and habitat descriptors. Correlation coefficients showed positive associations between species richness and each single environmental variable. Mean annual temperature was the strongest environmental predictor of raptor species richness (explaining 82% of the variance), consistent with the ambient energy hypothesis. Another descriptor of ambient energy (PET) explained 75% of the spatial variation. Both the AET and the vegetation structure coefficient explained 77% of the spatial variation in richness. The spatial clusters of extreme residuals identified the subtropical rainforests and the arid heights and low plateaux of the study area as regions where local environmental conditions appear to interfere with the general trend identified by the model at the regional scale.
Spatially explicit fisheries simulation models for policy evaluation
Authors: Pelletier Dominique , Mahévas Stéphanie
Published: November 2005

This paper deals with the design of modelling tools suitable for investigating the consequences of alternative policies on the dynamics of resources and fisheries, such as the evaluation of marine protected areas (MPA). We first review the numerous models that have been developed for this purpose, and compare them from several standpoints: population modelling, exploitation modelling and management measure modelling. We then present a generic fisheries simulation model, Integration of Spatial Information for FISHeries simulation (ISIS‐Fish). This spatially explicit model allows quantitative policy screening for fisheries with mixed‐species harvests. It may be used to investigate the effects of combined management scenarios including a variety of policies: total allowable catch (TAC), licenses, gear restrictions, MPA, etc. Fisher's response to management may be accounted for by means of decision rules conditioned on population and exploitation parameters. An application to a simple example illustrates the relevance of this kind of tool for policy screening, particularly in the case of mixed fisheries. Finally, the reviewed models and ISIS‐Fish are discussed and confronted in the light of the underlying assumptions and model objectives. In the light of this discussion, we identify desirable features for fisheries simulation models aimed at policy evaluation, and particularly MPA evaluation.
Wasted fishery resources: discarded by‐catch in the USA
Authors: Harrington Jennie M , Myers Ransom A , Rosenberg Andrew A
Published: November 2005

Fishery by‐catch, especially discarded by‐catch, is a serious problem in the world's oceans. Not only are the stocks of discarded species affected, but entire trophic webs and habitats may be disrupted at the ecosystem level. This paper reviews discarding in the marine fisheries of the USA; however, the type, diversity and regulatory mechanisms of the fisheries are similar to developed fisheries and management programmes throughout the world. We have compiled current estimates of discarded by‐catch for each major marine fishery in the USA using estimates from existing literature, both published and unpublished. We did not re‐estimate discards or discard rates from raw data, nor did we include data on protected species (turtles, mammals and birds) and so this study covers discarded by‐catch of finfish and fishable invertebrates. For some fisheries, additional calculations were required to transform number data into weight data, and typically length and weight composition data were used. Specific data for each fishery are referenced in Harrington et al. (Wasted Resources: Bycatch and discards in US Fisheries, Oceana, Washington, DC, 2005). Overall, our compiled estimates are that 1.06 million tonnes of fish were discarded and 3.7 million tonnes of fish were landed in USA marine fisheries in 2002. This amounts to a nationwide discard to landings ratio of 0.28, amongst the highest in the world. Regionally, the southeast had the largest discard to landings ratio (0.59), followed closely by the highly migratory species fisheries (0.52) and the northeast fisheries (0.49). The Alaskan and west coast fisheries had the lowest ratios (0.12 and 0.15 respectively). Shrimp fisheries in the southeast were the major contributors to the high discard rate in that region, with discard ratios of 4.56 (Gulf of Mexico) and 2.95 (South Atlantic). By‐catch and discarding is a major component of the impact of fisheries on marine ecosystems. There have been substantial efforts to reduce by‐catch in some fisheries, but broadly based programmes covering all fisheries are needed within the USA and around the world. In response to international agreements to improve fishery management, by‐catch and discard reduction must become a regular part of fishery management planning.
Ambivalent Sexism, Scriptural Literalism, and Religiosity
Authors: Burn Shawn Meghan , Busso Julia
Published: November 2005

This correlational study explores the hypothesis that religiosity and scriptural literalism (the degree to which one interprets scriptures literally) are associated with sexism. Participants were female and male ( N = 504) university students who anonymously completed the Ambivalent Sexism Inventory ( Glick & Fiske, 1996 , 1997 , 2001a , 2001b ), the Scriptural Literalism Scale ( Hogge & Friedman, 1967 ), and the Religious Orientation Scale-Revised ( Gorsuch & McPherson, 1989 ). Intrinsic religiosity, extrinsic religiosity, and scriptural literalism were positively associated with benevolent, but not hostile, sexism. Intrinsic religiosity and scriptural literalism were positively related to the protective paternalism subscale, whereas extrinsic religiosity was related to the heterosexual intimacy, complementary gender differentiation, and protective paternalism subscales.
Sun Protection Policies in Miami‐Dade County Public Schools: Opportunities for Skin Cancer Prevention
Authors: Kirsner Robert S. , Parker Dorothy F. , Brathwaite Noel , Thomas Andrea , Tejada Francisco , Trapido Edward J.
Published: November 2005

Abstract:  Childhood exposure to ultraviolet radiation from the sun and a history of sunburns are risk factors for skin cancer. Because children spend time outdoors when they are at school, school sun protection policies are an important health issue, particularly in areas of the country with year‐round warm and sunny climates, such as Florida. To better understand the sun protection policies and practices in South Florida schools, a sample (n = 51) of elementary and middle schools in Miami‐Dade County public schools were surveyed as part of a CDC‐funded cancer control program at the University of Miami. Of the principals and teachers surveyed, most (78%) knew about the county school system's guidelines for avoiding excessive heat exposure, which include two sun protection measures. Two‐thirds reported that they shared these guidelines with teachers; 21% shared them with parents. Few schools monitor implementation of the guidelines, although 70% schedule outdoor activities to avoid peak sun hours. No schools required sunscreen, hats, or protective clothing. Physical education teachers and students spend an average of 4.5 and 0.6 hours per day outdoors, respectively. Improved school sun protection policies and monitoring of such policies is needed to reduce sun exposure and skin cancer risk for both students and staff.
Atypical Pyoderma Gangrenosum as a Manifestation of Childhood Acute Lymphoblastic Leukemia
Authors: Bakhshi Sameer , Sethuraman Gomathy , Singh Manoj K. , Arya Laxman Singh
Published: November 2005

Abstract:  Pyoderma gangrenosum is a neutrophilic dermatosis that may occur idiopathically or in association with various systemic diseases and malignancy. Although the association of this entity with myeloid malignancies is well known, its association with lymphoid malignancy is extremely rare. We describe atypical pyoderma gangrenosum in association with acute lymphoblastic leukemia in a 2‐year‐old child, an occurrence not reported before.
Purpura after Application of EMLA Cream in Two Children
Authors: Neri Iria , Savoia Francesco , Guareschi Elisa , Medri Matelda , Patrizi Annalisa
Published: November 2005

Abstract:  The eutectic mixture of local anesthetic cream, a 1 : 1 mixture of prilocaine and lidocaine, 2.5% each, is frequently used in pediatric and dermatologic practice to obtain local anesthesia. Side effects include transient skin blanching, erythema, urticaria, allergic contact dermatitis, irritant contact dermatitis, hyperpigmentation, and purpura. We report two children with a purpuric reaction after application of this mixture cream. Purpura after application of this anesthetic cream is a rare nonallergic reaction and only 17 occurrences have been reported, to our knowledge, in the literature. Patch tests could not be performed in our two patients because of lack of parental consent but we suggest that the purpuric reactions were most probably of toxic origin. The pathogenesis of purpura after application of eutectic mixture of local anesthetics cream, which resolves within 2 weeks without dermatologic sequelae and without any specific therapy, is complex. The lesions are probably caused by the direct effect of the cream components on the vessels but many other factors, such as atopic dermatitis, prematurity, subjective predisposition to purpura, trauma, and thrombocytopenia may play important pathogenetic roles.
Design Considerations of Volute Geometry of a Centrifugal Blood Pump
Authors: Chan Weng Kong , Wong Yew Wah , Hu Wei
Published: November 2005

Abstract:  This article compares two different design techniques that are conventionally used in the design of volutes for centrifugal pumps. The imbalanced forces due to the geometry of the volute need to be taken into consideration especially in centrifugal blood pumps with magnetically suspended impeller. A reduction of these forces can reduce the instability of the impeller motion as well as the power needed to counteract its influence. Volutes using the constant angular momentum (CAM) and the constant mean velocity (CMV) methods were developed and modeled numerically. The computational results on the effect of volute geometry on the performance of a centrifugal blood pump impeller for six different volutes are presented here. For volutes designed using the CAM method, model B (volute expansion angle of 3°) had the lowest radial force of 0.26 N while the pressure head generated was 12 900 Pa. For volutes designed using the CMV method, model F (1.6 m/s) had the lowest imbalanced force of 0.45 N. However, the pressure developed by this pump was also one of the lowest at 10 652 Pa. Furthermore, when the peak scalar stresses and the mean exposure time of particles for all designs were determined using Lagrangian particle tracking method, it was observed that in general, the peak scalar stresses in CAM designed volutes are lower than those designed using CMV method. The mean exposure time of particles in the pump ranged from 400 to 500 ms. The simulation results showed that the volute designed using CAM method was superior to that of a CMV volute in terms of the magnitude of the radial force and the peak scalar stresses for the same pressure head generated. Results show that the design of volutes for blood pumps should go beyond conventional empirical methods to obtain optimal results.
Mechanism of Repolarization Alternans Has Restitution of Action Potential Duration Dependent and Independent Components
Authors: WU RUNZE , PATWARDHAN ABHIJIT
Published: November 2005

Introduction: Investigation of relationship between diastolic‐interval (DI)‐dependent restitution of action potential duration (APD) and alternans of APD has produced conflicting results. We used a novel pacing protocol to determine the role of restitution in alternans by minimizing changes in DI preceding each activation.

Methods: Transmembrane potentials were recorded from right ventricular endocardial tissue isolated from five dogs. We used three pacing sequences: (i) The tissue was paced at a constant DI for 100 beats. (ii) The DIs were changed randomly between two sequences of constant DI. (iii) Each constant DI trial was followed by constant cycle length trial where pacing cycle length was equal to average cycle length during previous constant DI trial.

Results: Alternans of APD occurred even when DIs preceding each activation were invariant. Slopes of restitution during constant DI pacing were both negative and positive and were much larger than unity. Alternans amplitude during constant cycle length pacing was larger than during constant DI, 32.2 ± 12.3 versus 7.5 ± 2.8 msec, P < 0.01. Random perturbation of DI decreased alternans amplitude during constant DI pacing from 14.7 ± 4.8 to 10.5 ± 3.4 msec, P < 0.01.

Conclusion: Our results indicate that mechanism of repolarization alternans has restitution‐dependent and restitution‐independent components. However, our results also provide direct evidence that shows that DI‐dependent restitution of APD is not a necessary mechanism for the alternans to exist. Ability to pace with explicit control of DI provides a novel approach to dissect mechanisms of alternans into restitution‐dependent and restitution‐independent effects.
Catheter Ablation of Ventricular Fibrillation Storm in Patients with Infiltrative Amyloidosis of the Heart
Authors: MLCOCHOVA HANKA , SALIBA WALID I. , BURKHARDT DAVID J. , RODRIGUEZ RENE E. , CUMMINGS JENNIFER E. , LAKKIREDDY DHANUNJAYA , PATEL DIMPI , NATALE ANDREA
Published: November 2005

Recent studies have demonstrated that premature ventricular contractions (PVCs) originating from the Purkinje system are responsible for initiation of ventricular fibrillation (VF) in patients with and without structural heart disease. Ablation of the PVCs has been shown to be feasible. We report 2 patients with repetitive VF associated with cardiac amyloidosis. Each episode of ventricular arrhythmia was preceded by monomorphic PVC. The electrical storms were drug resistant. Electrophysiological testing was performed and the sites of earliest activation were localized within the left ventricle in the absence of significant scar tissue. After ablation, PVCs subsided and there were no further VF recurrences.
Comparison of Radiofrequency Ablation in Normal Versus Scarred Myocardium
Authors: KOVOOR PRAMESH , DALY MICHAEL P. J. , POULIOPOULOS JIM , BYTH KAREN , DEWSNAP BARBARA I. , EIPPER VICKI E. , YUNG TERESA , UTHER JOHN F. B. , ROSS DAVID L.
Published: November 2005

Introduction: Reentrant circuits causing ventricular tachycardia are closely associated with previously scarred myocardium. The presence of scar has been blamed for the poor success rate of radiofrequency ablation (RFA) in that context. This article investigates the in vivo effects of radiofrequency ablation in myocardium scarred from acute myocardial infarction.

Methods and Results: Anterior myocardial infarction was induced in five dogs by ligating the left anterior descending artery. The mean left ventricular ejection fraction after infarction was 38%. At a mean of 15 weeks following myocardial infarction, 50 RFA lesions were created in random order, 25 in scarred and 25 in normal myocardium using a needle electrode (21 gauge, 5 mm in length) introduced from the epicardium of the left ventricle at thoracotomy. During unipolar temperature‐controlled RFA (90°C for 60 seconds), intramural temperatures were measured by thermistors at distances of 1, 2, 3, 4, and 5 mm from the ablating electrode. The margins of the lesions were clearly discernible in scar at histological examination in 64% of ablations where the scarring was patchy. There were no significant differences between lesion sizes, intramural temperatures at different distances, total energy required for ablation, or mean impedance during ablation of normal versus scarred myocardium.

Conclusions: Scar does not affect lesion size or intramural temperature profile during RFA if electrode size, tissue contact, and tip temperature are controlled. More radiofrequency energy is not required to maintain tip temperature at 90°C in scar compared to normal myocardium.
Proarrhythmia of Circumferential Left Atrial Lesions for Management of Atrial Fibrillation
Authors: DAOUD EMILE G. , WEISS RAUL , AUGOSTINI RALPH , HUMMEL JOHN D. , KALBFLEISCH STEVEN J. , VAN DEREN JOHN M. , DAWSON GEORGIA , BOWMAN KATHY
Published: November 2005

Background: After circumferential ablation for atrial fibrillation, new onset left atrial flutter (LA Flr) may occur. This study assessed the relationship between induced and clinical episodes of LA Flr, the rate of spontaneous resolution of LA Flr, and the proarrhythmic effect of circumferential ablation.

Methods and Results: A total 112 patients underwent circumferential LA ablation for atrial fibrillation. Immediately after completion of the ablation, LA Flr was induced in 43 of 112 (38%) patients, but was not targeted for ablation. During follow‐up (14 ± 4 months), new onset LA Flr occurred in 28 of 112 (25%) patients; however, the presence of inducible LA Flr did not identify those patients with clinical LA Flr (P = 0.6). In comparison to episodes of atrial fibrillation occurring before circumferential ablation, LA Flr was associated with a faster ventricular rate (124 ± 19 beats/min vs 91 ± 16 beats/min, P < 0.001), and was more likely to be persistent requiring cardioversion (86% vs 32%, P = 0.01). By ≥4 months postcircumferential ablation, clinical LA Flr resolved in 18 of 28 patients (64%). A second ablation procedure for LA Flr was performed in 9 of 10 patients. Of the 17 morphologies, 16 (94%) LA Flr circuits were successfully ablated.

Conclusions: (1) LA Flrs that are induced immediately after circumferential ablation for atrial fibrillation do not identify those patients who require a second ablation procedure for clinical LA Flr; (2) Since the majority of clinical LA Flrs spontaneously resolve, ablation of LA Flr should be postponed several months; and (3) new onset LA Flr after ablation for atrial fibrillation is likely a manifestation of the proarrhythmic effect of ablation lines in the LA.
Co‐Firing and Shrinkage Matching in Low‐ and Middle‐ Permittivity Dielectric Compositions for a Low‐Temperature Co‐Fired Ceramics System
Authors: Choi Young‐Jin , Park Jeong‐Hyun , Ko Won‐Jun , Hwang In‐Sung , Park Jae‐Hwan , Park Jae‐Gwan , Nahm Sahn
Published: November 2005

Co‐firing and shrinkage matching of low‐ and middle‐ permittivity dielectric compositions for the application to a low‐temperature co‐fired ceramics system was investigated. Several glass compositions were initially examined for the low‐temperature firing of middle‐K (K∼20) dielectrics, and their compositions were further modified for shrinkage matching between low‐K (K∼6) and middle‐K layers. By controlling the processing parameters such as the solid loading ratio and stacking sequence, the warpages between hetero‐layers could be minimized.
Effects of short‐term normobaric hypoxia on haematology, muscle phenotypes and physical performance in highly trained athletes
Authors: Basset Fabien A. , Joanisse Denis R. , Boivin Frédéric , St‐Onge Josée , Billaut François , Doré Jean , Chouinard Richard , Falgairette Guy , Richard Denis , Boulay Marcel R.
Published: November 2005

This study aimed to determine the impact of short‐term normobaric hypoxia on physiology and performance in highly trained athletes. Twelve (7 male and 5 female) athletes were randomly assigned into two groups and spent 8 h per night for two consecutive nights a week over 3 weeks under either short‐term normobaric hypoxia (simulating 3636 m altitude, inspired O2= 13%) or in normobaric normoxia in a single‐blind study. Following a 3 week washout period, athletes were then exposed to the other condition. Athletes were tested for maximal oxygen consumption and time to exhaustion on an electromagnetically braked cycle ergometer before and after each treatment in addition to being tested for anaerobic performance (Wingate test) on a modified Monark cycle ergometer. Blood samples were taken throughout the experiment and vastus lateralis muscle biopsies were taken before and after each treatment. Increases in red blood cell count, haematocrit, haemoglobin, platelet number and erythropoietin concentration were observed following short‐term normobaric hypoxia. Except for a modest decrease in phosphofructokinase activity following short‐term normobaric hypoxia, no changes were observed in muscle enzyme activities, buffer capacity, capillary density or morphology. No performance measures were changed following short‐term normobaric hypoxia or normobaric normoxia. Although short‐term normobaric hypoxia exposure increased levels of a number of haematological parameters, this was not associated with improved aerobic or anaerobic performance in highly trained athletes.
P2X purinoceptor subtypes on paraventricular nucleus neurones projecting to the rostral ventrolateral medulla in the rat
Authors: Cham Joo Lee , Owens Neil C. , Barden Julian. A. , Lawrence Andrew J. , Badoer Emilio
Published: November 2005

The rostral ventrolateral medulla (RVLM) is essential for the generation of sympathetic nerve activity. The RVLM receives a substantial innervation from the hypothalamic paraventricular nucleus (PVN). Activation of P2X purinoceptors via ATP has been shown to mediate fast excitatory synaptic neurotransmission. There is mounting evidence to suggest the presence of P2X purinoceptors in hypothalamic nuclei, including the PVN. In this study, we determined whether P2X1–P2X6 purinoceptor subtypes were present on PVN neurones that projected to the RVLM. Injection of the retrogradely transported tracer, rhodamine‐tagged microspheres, into the pressor region of the RVLM was used to identify the neurones in the PVN that innervated the RVLM. P2X1–P2X6 purinoceptors were detected by immunohistochemistry. Double‐labelled neurones were quantified and expressed as a proportion of the retrogradely labelled neurones. The proportions of double‐labelled neurones for each of the P2X purinoceptor subtypes varied, on average, from 14 to 29%. The P2X3 purinoceptor subtype was found to be the dominant purinoceptor subtype present on PVN neurones projecting to the RVLM. Additionally it was apparent that more than one P2X purinoceptor subtype was present on the PVN neurones projecting to the RVLM, since the sum of the average percentages of double‐labelled neurones for each P2X purinoceptor subtype exceeded 100%. These findings highlight the presence of the P2X1–P2X6 purinoceptors on PVN neurones projecting to the RVLM. The results suggest a potential role for ATP in the PVN in the regulation of sympathetic nerve activity.
Simultaneous Mutations in CA and Vif of Maedi-Visna Virus Cause Attenuated Replication in Macrophages and Reduced Infectivity In Vivo
Authors: Gudmundsson Bjarki , Jónsson Stefán Ragnar , Ólafsson Oddur , Agnarsdóttir Gudrún , Matthíasdóttir Sigrídur , Georgsson Gudmundur , Torsteinsdóttir Sigurbjorg , Svansson Vilhjálmur , Kristbjornsdóttir Helga Bryndís , Franzdóttir Sigrídur Rut , Andrésson Ólafur S. , Andrésdóttir Valgerdur
Published: November 2005

Maedi-visna virus (MVV) is a lentivirus of sheep sharing several key features with the primate lentiviruses. The virus causes slowly progressive diseases, mainly in the lungs and the central nervous system of sheep. Here, we investigate the molecular basis for the differential growth phenotypes of two MVV isolates. One of the isolates, KV1772, replicates well in a number of cell lines and is highly pathogenic in sheep. The second isolate, KS1, no longer grows on macrophages or causes disease. The two virus isolates differ by 129 nucleotide substitutions and two deletions of 3 and 15 nucleotides in the
env
gene. To determine the molecular nature of the lesions responsible for the restrictive growth phenotype, chimeric viruses were constructed and used to map the phenotype. An L120R mutation in the CA domain, together with a P205S mutation in Vif (but neither alone), could fully convert KV1772 to the restrictive growth phenotype. These results suggest a functional interaction between CA and Vif in MVV replication, a property that may relate to the innate antiretroviral defense mechanisms in sheep.
Vaccinia Virus Nicking-Joining Enzyme Is Encoded by K4L (VACWR035)
Authors: Eckert Dawn , Williams Ollie , Meseda Clement A. , Merchlinsky Michael
Published: November 2005

Vaccinia virus encodes an enzyme with DNA modifying activity that cleaves and inefficiently cross-links cruciformic DNA. This enzyme is contained within the virion, expressed at late times postinfection, and processes DNA in an energy-independent, Mg
2+
ion-independent manner. Viral nuclease activity was measured in extracts from cells infected with well-defined viral mutants. Since some viral extracts lacked nuclease activity, the gene encoding the activity was postulated to be one of the open reading frames absent in the viruses lacking activity. Inducible expression of each candidate open reading frame revealed that only the gene VACWR035, or K4L, was required for nuclease activity. A recombinant virus missing only the open reading frame for K4L lacked nuclease activity. Extracts from a recombinant virus expressing K4L linked to a FLAG polypeptide were able to cleave and cross-link cruciformic DNA. There were no significant differences between the virus lacking K4L and wild-type vaccinia virus WR with respect to infectivity, growth characteristics, or processing of viral replicative intermediate DNA, including both telomeric and cross-linked forms. Purification of the K4L FLAG polypeptide expressed in bacteria yielded protein containing nicking-joining activity, implying that K4L is the only vaccinia virus protein required for the nicking-joining enzymatic activity.
Chimeric Recombinant Human Metapneumoviruses with the Nucleoprotein or Phosphoprotein Open Reading Frame Replaced by That of Avian Metapneumovirus Exhibit Improved Growth In Vitro and Attenuation In Vivo
Authors: Pham Quynh N. , Biacchesi Stéphane , Skiadopoulos Mario H. , Murphy Brian R. , Collins Peter L. , Buchholz Ursula J.
Published: November 2005

Chimeric versions of recombinant human metapneumovirus (HMPV) were generated by replacing the nucleoprotein (N) or phosphoprotein (P) open reading frame with its counterpart from the closely related avian metapneumovirus (AMPV) subgroup C. In Vero cells, AMPV replicated to an approximately 100-fold-higher titer than HMPV. Surprisingly, the N and P chimeric viruses replicated to a peak titer that was 11- and 25-fold higher, respectively, than that of parental HMPV. The basis for this effect is not known but was not due to obvious changes in the efficiency of gene expression. AMPV and the N and P chimeras were evaluated for replication, immunogenicity, and protective efficacy in hamsters. AMPV was attenuated compared to HMPV in this mammalian host on day 5 postinfection, but not on day 3, and only in the nasal turbinates. In contrast, the N and P chimeras were reduced approximately 100-fold in both the upper and lower respiratory tract on day 3 postinfection, although there was little difference by day 5. The N and P chimeras induced a high level of neutralizing serum antibodies and protective efficacy against HMPV; AMPV was only weakly immunogenic and protective against HMPV challenge, reflecting antigenic differences. In African green monkeys immunized intranasally and intratracheally, the mean peak titer of the P chimera was reduced 100- and 1,000-fold in the upper and lower respiratory tracts, whereas the N chimera was reduced only 10-fold in the lower respiratory tract. Both chimeras were comparable to wild-type HMPV in immunogenicity and protective efficacy. Thus, the P chimera is a promising live HMPV vaccine candidate that paradoxically combines improved growth in vitro with attenuation in vivo.
Novel Caprine Adeno-Associated Virus (AAV) Capsid (AAV-Go.1) Is Closely Related to the Primate AAV-5 and Has Unique Tropism and Neutralization Properties
Authors: Arbetman Alejandra E. , Lochrie Michael , Zhou Shangzhen , Wellman Jennifer , Scallan Ciaran , Doroudchi Mohammad M. , Randlev Britta , Patarroyo-White Susannah , Liu Tongyao , Smith Peter , Lehmkuhl Howard , Hobbs Lea Ann , Pierce Glenn F. , Colosi Peter
Published: November 2005

Preexisting humoral immunity to adeno-associated virus (AAV) vectors may limit their clinical utility in gene delivery. We describe a novel caprine AAV (AAV-Go.1) capsid with unique biological properties. AAV-Go.1 capsid was cloned from goat-derived adenovirus preparations. Surprisingly, AAV-Go.1 capsid was 94% identical to the human AAV-5, with differences predicted to be largely on the surface and on or under the spike-like protrusions. In an in vitro neutralization assay using human immunoglobulin G (IgG) (intravenous immune globulin [IVIG]), AAV-Go.1 had higher resistance than AAV-5 (100-fold) and resistance similar to that of AAV-4 or AAV-8. In an in vivo model, SCID mice were pretreated with IVIG to generate normal human IgG plasma levels prior to the administration of AAV human factor IX vectors. Protein expression after intramuscular administration of AAV-Go.1 was unaffected in IVIG-pretreated mice, while it was reduced 5- and 10-fold after administration of AAV-1 and AAV-8, respectively. In contrast, protein expression after intravenous administration of AAV-Go.1 was reduced 7.1-fold, similar to the 3.8-fold reduction observed after AAV-8administration in IVIG-pretreated mice, and protein expression was essentially extinguished after AAV-2 administration in mice pretreated with much less IVIG (15-fold). AAV-Go.1 vectors also demonstrated a marked tropism for lung when administered intravenously in SCID mice. The pulmonary tropism and high neutralization resistance to human preexisting antibodies suggest novel therapeutic uses for AAV-Go.1 vectors, including targeting diseases such as cystic fibrosis. Nonprimate sources of AAVs may be useful to identify additional capsids with distinct tropisms and high resistance to neutralization by human preexisting antibodies.
Specific Binding of
Autographa californica M
Nucleopolyhedrovirus Occlusion-Derived Virus to Midgut Cells of
Heliothis virescens
Larvae Is Mediated by Products of
pif
Genes
Ac119
and
Ac022
but Not by
Ac115
Authors: Ohkawa Taro , Washburn Jan O. , Sitapara Ronika , Sid Eric , Volkman Loy E.
Published: November 2005

Per os infectivity factors PIF1 (Ac119) and PIF2 (Ac022), like P74, are essential for oral infection of lepidopteran larval hosts of
Autographa californica M
nucleopolyhedrovirus (Ac
M
NPV). Here we show that Ac115 also is a PIF (PIF3) and that, unlike PIF1 and PIF2, it does not mediate specific binding of Ac
M
NPV occlusion-derived virus (ODV) to midgut target cells. We used an improved in vivo fluorescence dequenching assay to compare binding, fusion, and competition among control Ac
M
NPV ODV and the ODVs of Ac
M
NPV PIF1, PIF2, and PIF3 deletion mutants. Our results showed that binding and fusion of PIF1 and PIF2 mutants, but not the PIF3 mutant, were both qualitatively and quantitatively different from those of control ODV. Unlike control and PIF3-deficient ODV, an excess of PIF1- or PIF2-deficient ODV failed to compete effectively with control ODV's binding to specific receptors on midgut epithelial cells. Moreover, the levels of PIF1- and PIF2-deficient ODV binding were depressed threefold compared to control levels. Binding, fusion, and competition by PIF3-deficient ODV, however, were all indistinguishable from those of control ODV. These results implicated PIF1 and PIF2 as ODV envelope attachment proteins that mediate specific binding to primary target cells within the midgut. In contrast, PIF3 mediates another unidentified, but critical, early event during primary infection.
Phylogenetic Analysis of the Main Neutralization and Hemagglutination Determinants of All Human Adenovirus Prototypes as a Basis for Molecular Classification and Taxonomy
Authors: Madisch Ijad , Harste Gabi , Pommer Heidi , Heim Albert
Published: November 2005

Human adenoviruses (HAdV) are responsible for a wide spectrum of diseases. The neutralization ε determinant (loops 1 and 2) and the hemagglutination γ determinant are relevant for the taxonomy of HAdV. Precise type identification of HAdV prototypes is crucial for detection of infection chains and epidemiology. ε and γ determinant sequences of all 51 HAdV were generated to propose molecular classification criteria. Phylogenetic analysis of ε determinant sequences demonstrated sufficient genetic divergence for molecular classification, with the exception of HAdV-15 and HAdV-29, which also cannot be differentiated by classical cross-neutralization. Precise sequence divergence criteria for typing (<2.5% from loop 2 prototype sequence and <2.4% from loop 1 sequence) were deduced from phylogenetic analysis. These criteria may also facilitate identification of new HAdV prototypes. Fiber knob (γ determinant) phylogeny indicated a two-step model of species evolution and multiple intraspecies recombination events in the origin of HAdV prototypes. HAdV-29 was identified as a recombination variant of HAdV-15 (ε determinant) and a speculative, not-yet-isolated HAdV prototype (γ determinant). Subanalysis of molecular evolution in hypervariable regions 1 to 6 of the ε determinant indicated different selective pressures in subclusters of species HAdV-D. Additionally, γ determinant phylogenetic analysis demonstrated that HAdV-8 did not cluster with -19 and -37 in spite of their having the same tissue tropism. The phylogeny of HAdV-E4 suggested origination by interspecies recombination between HAdV-B (hexon) and HAdV-C (fiber), as in simian adenovirus 25, indicating additional zoonotic transfer. In conclusion, molecular classification by systematic sequence analysis of immunogenic determinants yields new insights into HAdV phylogeny and evolution.
Unique Ligand Binding Sites on CXCR4 Probed by a Chemical Biology Approach: Implications for the Design of Selective Human Immunodeficiency Virus Type 1 Inhibitors
Authors: Choi Won-Tak , Tian Shaomin , Dong Chang-Zhi , Kumar Santosh , Liu Dongxiang , Madani Navid , An Jing , Sodroski Joseph G. , Huang Ziwei
Published: November 2005

The chemokine receptor CXCR4 plays an important role as the receptor for the normal physiological function of stromal cell-derived factor 1α (SDF-1α) and the coreceptor for the entry of human immunodeficiency virus type 1 (HIV-1) into the cell. In a recent work (S. Tian et al., J. Virol.79:12667-12673, 2005), we found that many residues throughout CXCR4 transmembrane (TM) and extracellular loop 2 domains are specifically involved in interaction with HIV-1 gp120, as most of these sites did not play a role in either SDF-1α binding or signaling. These results provided direct experimental evidence for the distinct functional sites on CXCR4 for HIV-1 and the normal ligand SDF-1α. To further understand the CXCR4-ligand interaction and to develop new CXCR4 inhibitors to block HIV-1 entry, we have recently generated a new family of unnatural chemokines, termed synthetically and modularly modified (SMM) chemokines, derived from the native sequence of SDF-1α or viral macrophage inflammatory protein II (vMIP-II). These SMM chemokines contain various de novo-designed sequence replacements and substitutions byd-amino acids and display more enhanced CXCR4 selectivity, binding affinities, and/or anti-HIV activities than natural chemokines. Using these novel CXCR4-targeting SMM chemokines as receptor probes, we conducted ligand binding site mapping experiments on a panel of site-directed mutants of CXCR4. Here, we provide the first experimental evidence demonstrating that SMM chemokines interact with many residues on CXCR4 TM and extracellular domains that are important for HIV-1 entry, but not SDF-1α binding or signaling. The preferential overlapping in the CXCR4 binding residues of SMM chemokines with HIV-1 over SDF-1α illustrates a mechanism for the potent HIV-1 inhibition by these SMM chemokines. The discovery of distinct functional sites or conformational states influenced by these receptor sites mediating different functions of the natural ligand versus the viral or synthetic ligands has important implications for drug discovery, since the sites shared by SMM chemokines and HIV-1 but not by SDF-1α can be targeted for the development of selective HIV-1 inhibitors devoid of interference with normal SDF-1α function.
Baculovirus Entry into Human Hepatoma Cells
Authors: Matilainen Heli , Rinne Johanna , Gilbert Leona , Marjomäki Varpu , Reunanen Hilkka , Oker-Blom Christian
Published: November 2005

Autographa californica multiple nucleopolyhedrovirus
(AcMNPV), a prototype member of the
Baculoviridae
family, has gained increasing interest as a potential vector candidate for mammalian gene delivery applications. AcMNPV is known to enter both dividing and nondividing mammalian cell lines in vitro, but the mode and kinetics of entry as well as the intracellular transport of the virus in mammalian cells is poorly understood. The general objective of this study was to characterize the entry steps of AcMNPV- and green fluorescent protein-displaying recombinant baculoviruses in human hepatoma cells. The viruses were found to bind and transduce the cell line efficiently, and electron microscopy studies revealed that virions were located on the cell surface in pits with an electron-dense coating resembling clathrin. In addition, virus particles were found in larger noncoated plasma membrane invaginations and in intracellular vesicles resembling macropinosomes. In double-labeling experiments, virus particles were detected by confocal microscopy in early endosomes at 30 min and in late endosomes starting at 45 min posttransduction. Viruses were also seen in structures specific for early endosomal as well as late endosomal/lysosomal markers by nanogold preembedding immunoelectron microscopy. No indication of viral entry into recycling endosomes or the Golgi complex was observed by confocal microscopy. In conclusion, these results suggest that AcMNPV enters mammalian cells via clathrin-mediated endocytosis and possibly via macropinocytosis. Thus, the data presented here should enable future design of baculovirus vectors suitable for more specific and enhanced delivery of genetic material into mammalian cells.
Attenuation of Simian Immunodeficiency Virus SIVmac239 Infection by Prophylactic Immunization with DNA and Recombinant Adenoviral Vaccine Vectors Expressing Gag
Authors: Casimiro Danilo R. , Wang Fubao , Schleif William A. , Liang Xiaoping , Zhang Zhi-Qiang , Tobery Timothy W. , Davies Mary-Ellen , McDermott Adrian B. , O'Connor David H. , Fridman Arthur , Bagchi Ansu , Tussey Lynda G. , Bett Andrew J. , Finnefrock Adam C. , Fu Tong-ming , Tang Aimin , Wilson Keith A. , Chen Minchun , Perry Helen C. , Heidecker Gwendolyn J. , Freed Daniel C. , Carella Anthony , Punt Kara S. , Sykes Kara J. , Huang Lingyi , Ausensi Virginia I. , Bachinsky Margaret , Sadasivan-Nair Usha , Watkins David I. , Emini Emilio A. , Shiver John W.
Published: November 2005

The prophylactic efficacy of DNA and replication-incompetent adenovirus serotype 5 (Ad5) vaccine vectors expressing simian immunodeficiency virus (SIV) Gag was examined in rhesus macaques using an SIVmac239 challenge. Cohorts of either Mamu-A*01(+) or Mamu-A*01(−) macaques were immunized with a DNA prime-Ad5 boost regimen; for comparison, a third cohort consisting of Mamu-A*01(+) monkeys was immunized using the Ad5 vector alone for both prime and boost. All animals, along with unvaccinated control cohorts of Mamu-A*01(+) and Mamu-A*01(−) macaques, were challenged intrarectally with SIVmac239. Viral loads were measured in both peripheral and lymphoid compartments. Only the DNA prime-Ad5-boosted Mamu-A*01(+) cohort exhibited a notable reduction in peak plasma viral load (sevenfold) as well as in early set-point viral burdens in both plasma and lymphoid tissues (10-fold) relative to those observed in the control monkeys sharing the same Mamu-A*01 allele. The degree of control in each animal correlated with the levels of Gag-specific immunity before virus challenge. However, virus control was short-lived, and indications of viral escape were evident as early as 6 months postinfection. The implications of these results in vaccine design and clinical testing are discussed.
Identification of a Functional Envelope Protein from the HERV-K Family of Human Endogenous Retroviruses
Authors: Dewannieux Marie , Blaise Sandra , Heidmann Thierry
Published: November 2005

Genome-wide screening of sequence databases for human endogenous retroviruses (HERVs) has led to the identification of 18 coding
env
genes, among which two—the
syncytin
genes—encode fusogenic ENV proteins possibly involved in placenta physiology. Here we show that a third ENV, originating from the most “recent” HERV-K(HML2) family, is functional. Immunofluorescence analysis of
env
-transduced cells demonstrates expression of the protein at the cell surface, and we show that the protein confers infectivity to simian immunodeficiency virus pseudotypes. Western blot analysis of the pseudotyped virions further discloses the expected specific cleavage of the ENV precursor protein. This functional ENV could play a role in the amplification—via infection of the germ line—of the HERV-K genomic copies, all the more as coding HERV-K
gag
and
pol
genes can similarly be found in the human genome, which could therefore generate infectious virions of a fully endogenous origin.