bluuebunny/crossref_metadata_2025 huggingface dataset:
Candidate Antigens for Q Fever Serodiagnosis Revealed by Immunoscreening of a
Coxiella burnetii
Protein Microarray
Authors: Beare Paul A. , Chen Chen , Bouman Timo , Pablo Jozelyn , Unal Berkay , Cockrell Diane C. , Brown Wendy C. , Barbian Kent D. , Porcella Stephen F. , Samuel James E. , Felgner Philip L. , Heinzen Robert A.
Published: October 2008

Q fever is a widespread zoonosis caused by
Coxiella burnetii
. Diagnosis of Q fever is usually based on serological testing of patient serum. The diagnostic antigen of test kits is formalin-fixed phase I and phase II organisms of the Nine Mile reference strain. Deficiencies of this antigen include (i) potential for cross-reactivity with other pathogens; (ii) an inability to distinguish between
C. burnetii
strains; and (iii) a need to propagate and purify
C. burnetii
, a difficult and potentially hazardous process. Consequently, there is a need for sensitive and specific serodiagnostic tests utilizing defined antigens, such as recombinant
C. burnetii
protein(s). Here we describe the use of a
C. burnetii
protein microarray to comprehensively identify immunodominant antigens recognized by antibody in the context of human
C. burnetii
infection or vaccination. Transcriptionally active PCR products corresponding to 1,988
C. burnetii
open reading frames (ORFs) were generated. Full-length proteins were successfully synthesized from 75% of the ORFs by using an
Escherichia coli
-based in vitro transcription and translation system (IVTT). Nitrocellulose microarrays were spotted with crude IVTT lysates and probed with sera from acute Q fever patients and individuals vaccinated with Q-Vax. Immune sera strongly reacted with approximately 50
C. burnetii
proteins, including previously identified immunogens, an ankyrin repeat-domain containing protein, and multiple hypothetical proteins. Recombinant protein corresponding to selected array-reactive antigens was generated, and the immunoreactivity was confirmed by enzyme-linked immunosorbent assay. This sensitive and high-throughput method for identifying immunoreactive
C. burnetii
proteins will aid in the development of Q fever serodiagnostic tests based on recombinant antigen.
Reliability of Rapid Subtyping Tools Compared to That of Phylogenetic Analysis for Characterization of Human Immunodeficiency Virus Type 1 Non-B Subtypes and Recombinant Forms
Authors: Holguín África , López Marisa , Soriano Vincent
Published: October 2008

Human immunodeficiency virus type 1 (HIV-1) subtyping is often estimated on the basis of
pol
sequences by using online websites instead of phylogenetic analysis (phy). We evaluated the reliability of distinct rapid subtyping tools versus phy with a large panel of HIV-1 non-B subtypes and circulating recombinant forms (CRF).
pol
sequences (277 protease [PR] and 171 reverse transcriptase [RT] sequences) previously assigned by phy to eight distinct HIV-1 non-B subtypes were obtained from 277 HIV-infected patients. Phy was run again to identify CRF. Subtyping was then performed using three rapid tools (the Stanford, NCBI, and REGA online tools). Thirty-three additional clade B sequences were tested as controls. New phylogenetic analyses reclassified two-thirds of
pol
sequences previously assigned to HIV-1 non-B clades as CRF. CRF02_AG variants were correctly assigned by the Stanford and NCBI tools for 92 to 97% and 96 to 99% of PR-RT sequences, respectively, while they were correctly assigned by the REGA tool for only 18 to 32% of PR-RT sequences. The Stanford, NCBI, and REGA tools failed to assign pure non-B clades correctly for 24 to 33%, 35%, and 57 to 64% of PR-RT sequences, respectively. For PR-RT sequences from CRF other than CRF02_AG, discrepancies occurred in 98 to 100%, 18 to 43%, and 80 to 87% of sequences, respectively. The concordance between those tools and phy was almost complete for subtype B assignment. Rapid subtyping tools show relatively low agreement with phy in identifying HIV-1 non-B clades and CRF other than CRF02_AG. The Stanford tool shows the best concordance with phy for the assignment of pure non-B clades, while the NCBI tool performs better at identifying CRF. Before entering routine clinical use, rapid subtyping tools should be optimized and updated periodically. Larger numbers of different non-B subtypes and CRF sequences should be included.
Detection of a Molecular Biomarker for Zygomycetes by Quantitative PCR Assays of Plasma, Bronchoalveolar Lavage, and Lung Tissue in a Rabbit Model of Experimental Pulmonary Zygomycosis
Authors: Kasai Miki , Harrington Susan M. , Francesconi Andrea , Petraitis Vidmantas , Petraitiene Ruta , Beveridge Mara G. , Knudsen Tena , Milanovich Jeffery , Cotton Margaret P. , Hughes Johanna , Schaufele Robert L. , Sein Tin , Bacher John , Murray Patrick R. , Kontoyiannis Dimitrios P. , Walsh Thomas J.
Published: October 2008

We developed two real-time quantitative PCR (qPCR) assays, targeting the 28S rRNA gene, for the diagnosis of zygomycosis caused by the most common, clinically significant Zygomycetes. The amplicons of the first qPCR assay (qPCR-1) from
Rhizopus, Mucor
, and
Rhizomucor
species were distinguished through melt curve analysis. The second qPCR assay (qPCR-2) detected
Cunninghamella
species using a different primer/probe set. For both assays, the analytic sensitivity for the detection of hyphal elements from germinating sporangiospores in bronchoalveolar lavage (BAL) fluid and lung tissue homogenates from rabbits was 1 to 10 sporangiospores/ml. Four unique and clinically applicable models of invasive pulmonary zygomycosis served as surrogates of human infections, facilitating the validation of these assays for potential diagnostic utility. For qPCR-1, 5 of 98 infarcted lung specimens were positive by qPCR and negative by quantitative culture (qCx). None were qCx positive only. Among 23 BAL fluid samples, all were positive by qPCR, while 22 were positive by qCx. qPCR-1 detected
Rhizopus
and
Mucor
DNA in 20 (39%) of 51 serial plasma samples as early as day 1 postinoculation. Similar properties were observed for qPCR-2, which showed greater sensitivity than qCx for BAL fluid (100% versus 67%;
P
= 0.04;
n
= 15). The assay detected
Cunninghamella
DNA in 18 (58%) of 31 serial plasma samples as early as day 1 postinoculation. These qPCR assays are sensitive and specific for the detection of
Rhizopus, Mucor, Rhizomucor
, and
Cunninghamella
species and can be used for the study and detection of infections caused by these life-threatening pathogens.
High Genetic Diversity of Nontypeable
Haemophilus influenzae
Isolates from Two Children Attending a Day Care Center
Authors: LaCross Nathan C. , Marrs Carl F. , Patel Mayuri , Sandstedt Sara A. , Gilsdorf Janet R.
Published: October 2008

Twenty-one nontypeable
Haemophilus influenzae
(NTHi) isolates from the throats of two healthy children were genotyped by multilocus sequence typing. Nine unique sequence types (STs) were identified. These STs were scattered throughout the phylogenetic tree of reported NTHi STs, demonstrating the high level of NTHi diversity found in colonized children.
Three-Class-Resistant Human Immunodeficiency Virus Type 1 Variant in a Drug-Naive Heterosexual Couple
Authors: Razzolini Francesca , Saladini Francesco , Malincarne Lisa , Vicenti Ilaria , Palladino Nicola , Romano Laura , Francisci Daniela , Zazzi Maurizio
Published: October 2008

Combination antiretroviral treatment was initiated in a heterosexual couple newly diagnosed with human immunodeficiency virus type 1 infection. Multiple genotypic drug resistance testing following early rebound of viral load revealed that the same three-class-resistant human immunodeficiency virus type 1 strain had been present in both patients since before initiation of treatment.
Self-Assembly of the Recombinant Capsid Protein of a Swine Norovirus into Virus-Like Particles and Evaluation of Monoclonal Antibodies Cross-Reactive with a Human Strain from Genogroup II
Authors: Almanza Horacio , Cubillos Carolina , Angulo Iván , Mateos Francisco , Castón José R. , van der Poel Wim H. M. , Vinje Jan , Bárcena Juan , Mena Ignacio
Published: October 2008

Noroviruses (NoVs) are responsible for the majority of gastroenteritis outbreaks in humans. Recently, NoV strains which are genetically closely related to human genogroup II (GII) NoVs have been detected in fecal specimens from swine. These findings have raised concern about the possible role of pigs as reservoirs for NoVs that could infect humans. To better understand the epidemiology of swine NoVs in both the swine and the human populations, rapid immunoassays are needed. In this study, baculovirus recombinants were generated to express the capsid gene of a swine NoV GII genotype 11 (GII.11) strain which self-assembled into virus-like particles (VLPs). Subsequently, the purified VLPs were used to evoke monoclonal antibodies (MAbs) in mice. A panel of eight promising MAbs was obtained and evaluated for their ability to bind to heterologous VLPs, denaturated antigens, and truncated capsid proteins. The MAbs could be classified into two groups: two MAbs that recognized linear epitopes located at the amino-terminal half (shell domain) of the swine NoV GII.11 VLPs and that cross-reacted with human GII.4 NoV VLPs. The other six MAbs bound to conformational epitopes and did not cross-react with the human GII.4 VLPs. To our knowledge, this is the first report on the characterization of MAbs against swine NoVs. The swine NoV VLPs and the MAbs described here may be further used for the design of diagnostic reagents that could help increase our knowledge of the prevalence of NoV infections in pigs and the possible role of pigs as reservoirs for NoVs.
Cross-Sectional Study of Nasopharyngeal Carriage of
Streptococcus pneumoniae
in Human Immunodeficiency Virus-Infected Adults in the Conjugate Vaccine Era
Authors: Onwubiko Chinwendu , Swiatlo Edwin , McDaniel Larry S.
Published: October 2008

Human immunodeficiency virus (HIV)-infected patients have an increased rate of pneumococcal infections. Within the HIV-infected population, patients with low CD4
+
cell counts have a higher rate of pneumococcal infection. The purpose of our study was to determine pneumococcal carriage and to examine the serotypes carried by HIV-infected patients after the introduction of the conjugate vaccine. Nasopharyngeal swabs were obtained from patients during routine clinic visits. Samples were cultured on blood agar plates with gentamicin and screened for alpha-hemolysis, optochin sensitivity, and bile solubility. Capsular serotypes were determined by multiplex PCR, multibead assay, or latex agglutination. Antibiotic susceptibility was determined by the Etest method. Multilocus sequence typing was also performed. Of the 175 patients enrolled, 120 patients had absolute CD4
+
cell counts above 200/mm
3
and 55 had counts below 200/mm
3
. A total of six (3.4%) patients carried pneumococci. All but one of these patients had received the 23-valent pneumococcal vaccine within the previous 5 years. Five of the isolates were serotypes that are not included in the 7-valent conjugate vaccine. Immunization with the pneumococcal polysaccharide vaccine does not prevent colonization in HIV-infected patients; however, the observation of carriage of serotypes not included in the conjugate vaccine may be due to herd immunity and serotype replacement effects in the general population.
Emergence of Amantadine-Resistant H3N2 Avian Influenza A Virus in South Korea
Authors: Lee Jiyeon , Song Young Jun , Park Jeung Hyun , Lee Jun-Han , Baek Yun Hee , Song Min-Suk , Oh Taek-Kyu , Han Heon-Seok , Pascua Philippe Noriel Q. , Choi Young-Ki
Published: October 2008

We found a relatively high frequency of unique amantadine-resistant H3N2 and H9N2 avian influenza viruses (Val27Ile on M2 protein) isolated from live poultry markets in South Korea and confirmed that a Val27Ile single substitution in the M2 protein is enough to acquire the amantadine resistance phenotype by using reverse-genetically created human-avian reassortant viruses.
Enhancement by Tumor Necrosis Factor Alpha of Dengue Virus-Induced Endothelial Cell Production of Reactive Nitrogen and Oxygen Species Is Key to Hemorrhage Development
Authors: Yen Yu-Ting , Chen Hseun-Chin , Lin Yang-Ding , Shieh Chi-Chang , Wu-Hsieh Betty A.
Published: October 2008

Hemorrhage is a severe manifestation of dengue disease. Virus strain and host immune response have been implicated as the risk factors for hemorrhage development. To delineate the complex interplay between the virus and the host, we established a dengue hemorrhage model in immune-competent mice. Mice inoculated intradermally with dengue virus develop hemorrhage within 3 days. In the present study, we showed by the presence of NS1 antigen and viral nuclei acid that dengue virus actively infects the endothelium at 12 h and 24 h after inoculation. Temporal studies showed that beginning at day 2, there was macrophage infiltration into the vicinity of the endothelium, increased tumor necrosis factor alpha (TNF-α) production, and endothelial cell apoptosis in the tissues. In the meantime, endothelial cells in the hemorrhage tissues expressed inducible nitric oxide synthase (iNOS) and nitrotyrosine. In vitro studies showed that primary mouse and human endothelial cells were productively infected by dengue virus. Infection by dengue virus induced endothelial cell production of reactive nitrogen and oxygen species and apoptotic cell death, which was greatly enhanced by TNF-α.NG-Nitro-l-arginine methyl ester andN-acetyl cysteine reversed the effects of dengue virus and TNF-α on endothelial cells. Importantly, hemorrhage development and the severity of hemorrhage were greatly reduced in mice lacking iNOS or p47phoxor treatment with oxidase inhibitor, pointing to the critical roles of reactive nitrogen and oxygen species in dengue hemorrhage.
RAG2
−/−
γ
c
−/−
Mice Transplanted with CD34
+
Cells from Human Cord Blood Show Low Levels of Intestinal Engraftment and Are Resistant to Rectal Transmission of Human Immunodeficiency Virus
Authors: Hofer Ursula , Baenziger Stefan , Heikenwalder Mathias , Schlaepfer Erika , Gehre Nadine , Regenass Stephan , Brunner Thomas , Speck Roberto F.
Published: October 2008

Rectal transmission is one of the main routes of infection by human immunodeficiency virus type 1 (HIV-1). To efficiently study transmission mechanisms and prevention strategies, a small animal model permissive for rectal transmission of HIV is mandatory. We tested the susceptibility of RAG2
−/−
γ
c
−/−
mice transplanted with human cord blood hematopoietic stem cells to rectal infection with HIV. We rectally exposed these humanized mice to cell-free and cell-associated HIV. All mice remained HIV negative as assessed by plasma viral load. The same mice infected intraperitoneally showed high levels of HIV replication. In the gut-associated lymphatic tissue, we found disproportionately smaller numbers of human cells than in other lymphoid organs. This finding may explain the observed resistance to rectal transmission of HIV. To increase the numbers of local HIV target cells and the likelihood of HIV transmission, we treated mice with different proinflammatory stimuli: local application of interleukin-1β, addition of seminal plasma to the inoculum, or induction of colitis with dextran sodium sulfate. These procedures attracted some human leukocytes, but the transmission rate was still very low. The humanized mice showed low levels of human engraftment in the intestinal tract and seem to be resistant to rectal transmission of HIV, and thus they are an unsuitable model for this application.
Persistent Gammaherpesvirus Replication and Dynamic Interaction with the Host In Vivo
Authors: Hwang Seungmin , Wu Ting-Ting , Tong Leming M. , Kim Kyeong Seon , Martinez-Guzman DeeAnn , Colantonio Arnaud D. , Uittenbogaart Christel H. , Sun Ren
Published: October 2008

Gammaherpesviruses establish life-long persistency inside the host and cause various diseases during their persistent infection. However, the systemic interaction between the virus and host in vivo has not been studied in individual hosts continuously, although such information can be crucial to control the persistent infection of the gammaherpesviruses. For the noninvasive and continuous monitoring of the interaction between gammaherpesvirus and the host, a recombinant murine gammaherpesvirus 68 (MHV-68, a gammaherpesvirus 68) was constructed to express a firefly luciferase gene driven by the viral M3 promoter (M3FL). Real-time monitoring of M3FL infection revealed novel sites of viral replication, such as salivary glands, as well as acute replication in the nose and the lung and progression to the spleen. Continuous monitoring of M3FL infection in individual mice demonstrated the various kinetics of transition to different organs and local clearance, rather than systemically synchronized clearance. Moreover, in vivo spontaneous reactivation of M3FL from latency was detected after the initial clearance of acute infection and can be induced upon treatment with either a proteasome inhibitor Velcade or an immunosuppressant cyclosporine A. Taken together, our results demonstrate that the in vivo replication and reactivation of gammaherpesvirus are dynamically controlled by the locally defined interaction between the virus and the host immune system and that bioluminescence imaging can be successfully used for the real-time monitoring of this dynamic interaction of MHV-68 with its host in vivo.
Biogenesis of Cytoplasmic Membranous Vesicles for Plant Potyvirus Replication Occurs at Endoplasmic Reticulum Exit Sites in a COPI- and COPII-Dependent Manner
Authors: Wei Taiyun , Wang Aiming
Published: October 2008

Single-stranded positive-sense RNA viruses induce the biogenesis of cytoplasmic membranous vesicles, where viral replication takes place. However, the mechanism underlying this characteristic vesicular proliferation remains poorly understood. Previously, a 6-kDa potyvirus membrane protein (6K) was shown to interact with the endoplasmic reticulum (ER) and to induce the formation of the membranous vesicles. In this study, the involvement of the early secretory pathway in the formation of the 6K-induced vesicles was investigated in planta. By means of live-cell imaging, it was found that the 6K protein was predominantly colocalized with Sar1, Sec23, and Sec24, which are known markers of ER exit sites (ERES). The localization of 6K at ERES was prevented by the coexpression of a dominant-negative mutant of Sar1 that disables the COPII activity or by the coexpression of a mutant of Arf1 that disrupts the COPI complex. The secretion of a soluble secretory marker targeting the apoplast was arrested at the level of the ER in cells overexpressing 6K or infected by a potyvirus. This blockage of protein trafficking out of the ER by 6K and the distribution of 6K toward the ERES may account for the aggregation of the 6K-bound vesicles. Finally, virus infection was reduced when the accumulation of 6K at ERES was inhibited by impairing either the COPI or COPII complex. Taken together, these results imply that the cellular COPI and COPII coating machineries are involved in the biogenesis of the potyvirus 6K vesicles at the ERES for viral-genome replication.
Llama Antibody Fragments with Cross-Subtype Human Immunodeficiency Virus Type 1 (HIV-1)-Neutralizing Properties and High Affinity for HIV-1 gp120
Authors: Forsman Anna , Beirnaert Els , Aasa-Chapman Marlén M. I. , Hoorelbeke Bart , Hijazi Karolin , Koh Willie , Tack Vanessa , Szynol Agnieszka , Kelly Charles , McKnight Áine , Verrips Theo , Haard Hans de , Weiss Robin A.
Published: October 2008

Members of theCamelidaefamily produce immunoglobulins devoid of light chains. We have characterized variable domains of these heavy chain antibodies, the VHH, from llamas immunized with human immunodeficiency virus type 1 (HIV-1) envelope protein gp120 in order to identify VHH that can inhibit HIV-1 infection. To increase the chances of isolating neutralizing VHH, we employed a functional selection approach, involving panning of phage libraries expressing the VHH repertoire on recombinant gp120, followed by a competitive elution with soluble CD4. By immunizing with gp120 derived from an HIV-1 subtype B′/C primary isolate, followed by panning on gp120 from HIV-1 isolates of subtypes A, B, and C, we could select for VHH with cross-subtype neutralizing activity. Three VHH able to neutralize HIV-1 primary isolates of subtypes B and C were characterized. These bound to recombinant gp120 with affinities close to the suggested affinity ceiling for in vivo-maturated antibodies and competed with soluble CD4 for this binding, indicating that their mechanism of neutralization involves interacting with the functional envelope spike prior to binding to CD4. The most potent VHH in terms of low 50% inhibitory concentration (IC50) and IC90values and cross-subtype reactivity was A12. These results indicate that camelid VHH can be potent HIV-1 entry inhibitors. Since VHH are stable and can be produced at a relatively low cost, they may be considered for applications such as HIV-1 microbicide development. Antienvelope VHH might also prove useful in defining neutralizing and nonneutralizing epitopes on HIV-1 envelope proteins, with implications for HIV-1 vaccine design.
Estimation of the Effective Number of Founders That Initiate an Infection after Aphid Transmission of a Multipartite Plant Virus
Authors: Betancourt Mónica , Fereres Alberto , Fraile Aurora , García-Arenal Fernando
Published: October 2008

The fecundity of RNA viruses can be very high. Thus, it is often assumed that viruses have large populations, and RNA virus evolution has been mostly explained using purely deterministic models. However, population bottlenecks during the virus life cycle could result in effective population numbers being much smaller than reported censuses, and random genetic drift could be important in virus evolution. A step at which population bottlenecks may be severe is host-to-host transmission. We report here an estimate of the size of the population that starts a new infection when
Cucumber mosaic virus
(CMV) is transmitted by the aphid
Aphis gossypii
, based on the segregation of two CMV genotypes in plants infected by aphids that acquired the virus from plants infected by both genotypes. Results show very small effective numbers of founders, between one and two, both in experiments in which the three-partite genome of CMV was aphid transmitted and in experiments in which a fourth RNA, CMV satellite RNA, was also transmitted. These numbers are very similar to those published for
Potato virus Y
, which has a monopartite genome and is transmitted by aphids according to a different mechanism than CMV. Thus, the number of genomic segments seems not to be a major determinant of the effective number of founders. Also, our results suggest that the occurrence of severe bottlenecks during horizontal transmission is general for viruses nonpersistently transmitted by aphids, indicating that random genetic drift should be considered when modeling virus evolution.
Identification of Gammaretroviruses Constitutively Released from Cell Lines Used for Human Immunodeficiency Virus Research
Authors: Takeuchi Yasuhiro , McClure Myra O. , Pizzato Massimo
Published: October 2008

Three human cell lines used in human immunodeficiency virus research were found to be contaminated with previously undetected retroviruses. On the bases of partial nucleotide sequence, capsid protein antigenicity, vector mobilization, and receptor usage studies, these contaminants were shown to be replication competent and to belong to theGammaretrovirusgenus. While the TZM-bl cells harbor ecotropic murine leukemia virus (MLV), Jurkat J6 cells were found to release xenotropic MLV and the A3.01/F7 cells to produce gibbon ape leukemia virus. These findings highlight the importance of routine testing of cell lines for retrovirus contamination to prevent potential experimental artifacts and allow correct biohazard assessment.
British American Tobacco’s failure in Turkey
Authors: Lawrence S
Published: October 2008

Background and objectives:

Transnational tobacco companies (TTCs) considered Turkey an important, potential investment market because of its high consumption rates and domestic commitment to tobacco. This paper outlines how British American Tobacco (BAT) attempted to establish a joint venture with the government monopoly TEKEL, while waiting for privatisation and a private tender.

Methods:

Analysis of tobacco industry documents from the Guildford Depository and online tobacco document sources.

Results:

BAT failed to establish a market share in Turkey until 2000 despite repeated attempts to form a joint venture with Turkey’s tobacco monopoly, TEKEL, once the market liberalised in the mid 1980s.

Conclusions:

BAT’s failure in the Turkish market was due to a misguided investment strategy focused solely on acquiring TEKEL and is contrasted with Philip Morris success in Turkey despite both TTCs working within Turkey’s unstable and corrupt investing climate.
An algorithm for tailoring pharmacotherapy for smoking cessation: results from a Delphi panel of international experts
Authors: Bader P , McDonald P , Selby P
Published: October 2008

Background:

Evidence-based smoking cessation guidelines recommend nicotine replacement therapy (NRT), bupropion SR and varenicline as first-line therapy in combination with behavioural interventions. However, there are limited data to guide clinicians in recommending one form over another, using combinations, or matching individual smokers to particular forms.

Objective:

To develop decision rules for clinicians to guide differential prescribing practices and tailoring of pharmacotherapy for smoking cessation.

Methods:

A Delphi approach was used to build consensus among a panel of 37 international experts from various health disciplines. Through an iterative process, panellists responded to three rounds of questionnaires. Participants identified and ranked “best practices” used by them to tailor pharmacotherapy to aid smoking cessation. An independent panel of 10 experts provided cross-validation of findings.

Results:

There was a 100% response rate to all three rounds. A high level of consensus was achieved in determining the most important priorities: (1) factors to consider in prescribing pharmacotherapy: evidence, patient preference, patient experience; (2) combinations based on: failed attempt with monotherapy, patients with breakthrough cravings, level of tobacco dependence; (3) specific combinations, main categories: (a) two or more forms of NRT, (b) bupropion + form of NRT; (4) specific combinations, subcategories: (1a) patch + gum, (1b) patch + inhaler, (1c) patch + lozenge; (2a) bupropion + patch, (2b) bupropion + gum; (5) impact of comorbidities on selection of pharmacotherapy: contraindications, specific pharmacotherapy useful for certain comorbidities, dual purpose medications; (6) frequency of monitoring determined by patient needs and type of pharmacotherapy.

Conclusion:

An algorithm and guide were developed to assist clinicians in prescribing pharmacotherapy for smoking cessation. There appears to be good justification for “off-label” use such as higher doses of NRT or combination therapy in certain circumstances. This practical tool reflects best evidence to date of experts in tobacco cessation.
Freezing resistance of gametophytes of the temperate fern, Polystichum retroso-paleaceum
Authors: Sato T. , Sakai A.
Published: October 2008

To understand the adaptation of the life cycle of ferns to cold regions, the freezing resistance of gametophytes and sporophytes of the temperate fern, Polystichum retroso-paleaceum, was evaluated in Sapporo. The mature sporophytes were hardy to −15 °C, but not to −20 °C. However, marginal cells located in the wing part of the gametophytes remained alive after freezing to −70 °C, and even to −196 °C. It was also observed that these surviving cells, after freezing to very low temperatures, resumed growth and grew into new gametophytes. Present evidence provides a demonstration of the adaptability to cold climates of the gametophytic generation of ferns.
Hyperparasitism of Scytalidium uredinicola on western gall rust, Endocronartium harknessii
Authors: Tsuneda A. , Hiratsuka Y. , Maruyama P. J.
Published: October 2008

Scytalidium uredinicola is a destructive hyperparasite of western gall rust, Endocronartium harknessii, that reduces the inoculum potential of the rust. It disintegrates spores and the basal cell region of the active rust sori and produces abundant arthroconidia among the rust spores to give the infected sori a yellowish-green to whitish-gray appearance, depending on the stage of infection. Hyphae of S. uredinicola are able to penetrate the wood tissue of the galls and destroy rust hyphae to a depth of 300 μm below the sori. When the parasite hyphae accumulate in the wood tissue, rust hyphae below the invaded area also appear to be degraded. On water agar, hyphae of S. uredinicola disintegrate rust spores within 48–72 h, without penetration. The use of S. uredinicola for biological control of pine stem rusts is considered promising.
Trophic upsurge and the relationship between phytoplankton biomass and productivity in Smallwood Reservoir, Canada
Authors: Ostrofsky M. L. , Duthie H. C.
Published: October 2008

The primary productivities of a natural lake, a new reservoir, and an established reservoir were investigated in western Labrador over two seasons. Productivity of the new reservoir was about twice those of the other two sites. Possible causes for this increased productivity were examined. It was found that differences in productivity were due to differences in standing stocks of plankters only, and not due to differing photosynthetic efficiencies, better light, or a combination of these factors. Since changes in biomass may be predicted from changes in phosphorus concentration, these findings support the hypothesis that the increased productivity in new impoundments may be explained by the increase in phosphorus concentration.
The Southern Indian Lake Impoundment and Churchill River Diversion
Authors: Newbury R. W. , McCullough G. K. , Hecky R. E.
Published: October 2008

The 242 000-km2 Churchill River basin extends across the northern half of Alberta, Saskatchewan, and Manitoba. In 1976, hydraulic control structures were completed to divert 75% of the natural river flow of 958 m3∙s−1 across the drainage divide separating the Churchill and Nelson river basins in northern Manitoba. The diversion flows follow 300 km of tributary valleys southward to the Nelson River channel where a 30-yr, 10 000 MW hydroelectric scheme is being developed. The diversion was accomplished by damming the northern outlet of Southern Indian Lake, a 1977-km2 riverine lake on the Churchill channel (latitude 57°N, longitude 99°W). The dam caused a 3-m impoundment above the historical lake levels, which flooded 414 km2 of the backshore zone. Permafrost, or permanently frozen ground, is widespread in the uplands surrounding the lake. As bedrock occurred on only 14% of the postimpoundment shoreline, severe erosion of the frozen backshore deposits is now underway. A long period of instability is anticipated on lake shorelines and in river valleys affected by the altered hydraulic regime. Although the whole-lake water exchange time was increased by only 41% by the impoundment, the circulation patterns and exchange times in individual basins of the lake were changed dramatically when the Churchill waters were diverted at the southern end of the lake. The effects of the changing regimes on the aquatic habitats and fisheries of Southern Indian Lake have been investigated in pre- and post-impoundment studies undertaken by the Freshwater Institute of the Department of Fisheries and Oceans.
Primary Productivity of Southern Indian Lake before, during, and after Impoundment and Churchill River Diversion
Authors: Hecky R. E. , Guildford S. J.
Published: October 2008

The primary productivity of seven regions of Southern Indian Lake and neighboring Wood Lake was measured during open-water seasons from 1974 to 1978. The lake had regional differences in chlorophyll concentrations and daily rates of integral primary production in 1974 and 1975 prior to impoundment of the lake. Regions receiving Churchill River flow tended to have higher chlorophyll concentrations and production rates than those regions marginal to the flow. Impoundment of the lake resulted in higher efficiencies of primary production in all regions, as indicated by higher light-saturated rates of carbon uptake per unit chlorophyll and by higher initial slopes of the hyperbolic light response relation of the phytoplankton. Many large basins of the lake had light penetration reduced by high concentrations of suspended sediment from eroding shorelines, while other areas had relatively unchanged light penetration. The increased efficiency of carbon fixation per unit chlorophyll resulted in higher rates of integral production in those regions where light penetration was not greatly affected. Daily rates of integral primary production in lake regions where light penetration had decreased markedly were not significantly different after impoundment because efficiencies of light utilization were higher. Comparison of the mean water column light intensities for those turbid regions with the values of Ik(light intensity at the onset of light saturation) for phytoplankton indicated that these turbid regions are now light deficient on average. Phosphorus deficiency, as indicated by alkaline phosphatase activity per unit ATP, which was present before impoundment, has been eliminated as the mean water column light intensity declined below 5 mEinsteins∙m−2∙min−1. The light environment of a new reservoir can be a significant determinant of integral production, and predicting the consequences of impoundment on phytoplankton production requires accurate prediction of the light environment.
Comparison of Phosphorus Turnover Times in Northern Manitoba Reservoirs with Lakes of the Experimental Lakes Area
Authors: Planas Dolores , Hecky R. E.
Published: October 2008

The turnover time, τ, of [32P]PO4was measured during July and September in a natural lake and two lakes in northern Manitoba that were undergoing impoundment in 1976. The τ values were 10–1000 times longer than those observed in the epilimnia and hypolimnia of two lakes being experimentally enriched in northwestern Ontario (Experimental Lakes Area) in the same summer. Algal populations in the Experimental Lakes Area exhibited short τ values over a large range of temperature and light values, which exceeded the ranges of temperatures and light intensities measured in northern Manitoba. In northern Manitoba the shortest τ values occurred in July when an independent indicator of sestonic P deficiency, the particulate alkaline phosphatase activity per unit weight ATP, measured moderate to severe P deficiency. In September in northern Manitoba, τ values were at least 10 times greater than in July and were among the highest reported in the literature. The 2-yr-old Notigi Reservoir had the highest τ values in both July and September, and it deviated strongly from the expected chlorophyll–phosphorus relation for north temperate lakes. These new reservoirs in northern Manitoba are not P limited and even the natural lakes may be only moderately P limited.
Effect of Impoundment and River Diversion on Profundal Macrobenthos of Southern Indian Lake Manitoba
Authors: Wiens Allen P. , Rosenberg David M.
Published: October 2008

Profundal macrobenthos in Southern Indian Lake, Manitoba, were surveyed to determine effects of hydroelectric manipulations in 1972 (preimpoundment), 1977 (postimpoundment), and 1979 (postdiversion). Lakewide average standing crops increased following impoundment and decreased by 3 yr after impoundment. Regional changes in standing crops usually could be related to additions of nutrients leached from flooded vegetation, additions of particulate organic matter resulting from shoreline erosion, and changes in integral primary production and suspended solids concentrations before and after impoundment. Greater increases in standing crops of macrobenthos in shallower compared with deeper depth zones of the lake after flooding were attributed to preferential deposition of organic materials in the shallow areas of the lake. Mean standing crops of macrobenthos were higher in regions through which the Churchill River flowed than in regions isolated from the flow before and after impoundment. After diversion, greatest decreases in standing crops occurred in isolated regions, whereas those of regions in the flow declined much less or increased. Responses of the most abundant taxa of macrobenthos (Pontoporeia brevicornis grp., Chironomidae, Oligochaeta, Sphaeriidae) differed in many ways from those recorded for other new reservoirs. Pontoporeia brevicornis grp. remained the most abundant benthic organism, there was no evident succession of macrobenthic taxa, and a high diversity of profundal species was maintained. These results, together with the slight changes in standing crop observed after flooding, indicated only a marginal impact on macrobenthos caused by the low-level flooding of Southern Indian Lake.
Breakdown of Conifer Needle Debris in a New Northern Reservoir, Southern Indian Lake, Manitoba
Authors: Crawford Patrick J. , Rosenberg David M.
Published: October 2008

Formation of the Southern Indian Lake reservoir, northern Manitoba, added an estimated 5.4 × 105 t of Picea mariana (black spruce) needles to the lake. The breakdown of needles in the lake was measured by stringing needles on monofilament line, placing the strings into 3-mm mesh bags, and situating the bags along four shorelines representing high and low shoreline erosion rates (clay vs. bedrock) and wave exposures (highly exposed vs. protected). Sampling was done in phases of 0–41 and 328–384 d. Effects of excluding macroinvertebrates were tested by using needle strings placed in 50-μm mesh bags at the clay low exposure shoreline. Needle breakdown apparently occurred in two stages. Initially, weight losses were due primarily to leaching and microbial conditioning. Subsequently, weight losses were due primarily to feeding by macroinvertebrates. Processing coefficients (k) ranged from 0.0011∙d−1 for the 50-μm mesh bags to 0.0097∙d−1 for the 3-mm mesh bags at the bedrock high exposure shoreline. The presence of macroinvertebrates and exposure to waves enhanced needle breakdown. Macroinvertebrate colonization of needle strings occurred rapidly [Formula: see text] and was mainly by chironomid larvae. Phaenopsectra punctipes (Wied.) and Brillia flavifrons (joh.) larvae fed directly on needle tissue. Macroinvertebrates accounted for ~ 40% of total weight losses and, apparently, were active over the winter. Leaching and microbial conditioning each accounted for ~ 30% of total weight losses. Values of k for Southern Indian Lake most resembled those for conifer needle breakdown recorded in streams, indicating the significance of wave action in the lake. The breakdown of P. mariana needles in Southern Indian Lake appeared to be a significant source of carbon during the year following impoundment, and needles may have been an important habitat for macroinvertebrates within localized areas.
Descriptions of Two New Species of Tanypodinae (Diptera:Chironomidae) from Southern Indian Lake, Canada
Authors: Bilyj Bohdan
Published: October 2008

Adults, pupae, and larvae of Guttipelopia rosenbergi n.sp. and Larsia canadensis n.sp. are described with notes on their distributions, phenologies, and habitat preferences. Taxonomic comparisons are made with other known Nearctic species within the respective genera.
Response of a Boreal Northern Pike (Esox lucius) Population to Lake Impoundment: Wupaw Bay, Southern Indian Lake, Manitoba
Authors: Bodaly R. A. , Lesack L. F. W.
Published: October 2008

A population of northern pike (Esox lucius) in Wupaw Bay, Southern Indian Lake, Manitoba, was monitored for 1 yr prior to impoundment of the lake and for 5 yr after impoundment. Impoundment had a pronounced but transient effect on pike reproductive success. The abundance of young-of-the-year pike in 1977, the 1st yr following impoundment, was 4–10 times higher than in the following 3 yr. Flooded terrestrial vegetation was apparently conducive to pike reproductive success only in the 1st yr after inundation. A strong 1977 year-class was evident in gillnet catches 4 yr later. This year-class was slower growing and in poorer condition than other year-classes. The impoundment of Wupaw Bay had no discernible effect on the growth, condition, or mortality of the adult pike population, with the exception of changes ascribed to the presence of the large 1977 year-class. The growth of the adult pike population appeared to be limited by forage fish abundance, but variations in feeding on fish were largely unexplained and were probably due to natural variation. The year-to-year pattern of forage fish reproductive success could not be related to the timing of lake impoundment, but may have been related to summer water temperatures. Variations in feeding on invertebrates appeared to be due to postimpoundment increases in productivity, but these changes were insufficient to affect population parameters. Wupaw Bay surface area increased by only 9% as a result of lake impoundment, the smallest areal increase in the Southern Indian Lake reservoir. The influx of nutrients and flooded terrestrial vegetation was therefore much smaller than in other temperate reservoirs where increases in pike production have been observed.
Development and annotation of perennial Triticeae ESTs and SSR markers
Authors: Bushman B. Shaun , Larson Steve R. , Mott Ivan W. , Cliften Paul F. , Wang Richard R.-C. , Chatterton N. Jerry , Hernandez Alvaro G. , Ali Shahjahan , Kim Ryan W. , Thimmapuram Jyothi , Gong George , Liu Lei , Mikel Mark A.
Published: October 2008

Triticeae contains hundreds of species of both annual and perennial types. Although substantial genomic tools are available for annual Triticeae cereals such as wheat and barley, the perennial Triticeae lack sufficient genomic resources for genetic mapping or diversity research. To increase the amount of sequence information available in the perennial Triticeae, three expressed sequence tag (EST) libraries were developed and annotated for Pseudoroegneria spicata , a mixture of both Elymus wawawaiensis and E. lanceolatus , and a Leymus cinereus  × L. triticoides interspecific hybrid. The ESTs were combined into unigene sets of 8 780 unigenes for P. spicata, 11 281 unigenes for Leymus, and 7 212 unigenes for Elymus. Unigenes were annotated based on putative orthology to genes from rice, wheat, barley, other Poaceae, Arabidopsis, and the non-redundant database of the NCBI. Simple sequence repeat (SSR) markers were developed, tested for amplification and polymorphism, and aligned to the rice genome. Leymus EST markers homologous to rice chromosome 2 genes were syntenous on Leymus homeologous groups 6a and 6b (previously 1b), demonstrating promise for in silico comparative mapping. All ESTs and SSR markers are available on an EST information management and annotation database ( http://titan.biotec.uiuc.edu/triticeae/ ).
Changes of cytosine methylation induced by wide hybridization and allopolyploidy inCucumis
Authors: Chen Longzheng , Chen Jinfeng
Published: October 2008

We previously demonstrated that allopolyploidization could induce phenotypic variations and genome changes in a newly synthesized allotetraploid in Cucumis . To explore the molecular involvement of epigenetic phenomena, we investigated cytosine methylation in Cucumis by using methylation-sensitive amplified polymorphism (MSAP). Results revealed a twofold difference in the level of cytosine methylation between the reciprocal F1hybrids and the allotetraploid. Analysis of the methylation pattern indicated that methylation changed at 2.0% to 6.4% of total sites in both the F1hybrids and the allotetraploid compared with their corresponding parents. Furthermore, 68.2% to 80.0% of the changed sites showed an increase in cytosine methylation and a majority of the methylated sites were from the maternal parent. Observations in different generations of the allotetraploid found that the extent of change in cytosine methylation pattern between the S1and S2was significantly higher than that between the S2and S3, suggesting stability in advanced generations. Analysis of 7 altered sequences indicated their similarity to known functional genes or genes involved in regulating gene expression. Reverse transcription – polymerase chain reaction analysis suggested that at least two of the methylation changes might be related to gene expression changes, which further supports the hypothesis that DNA methylation plays a significant role in allopolyploidization.
Gene pool variation and phylogenetic relationships of an indigenous northeast Italian grapevine collection revealed by nuclear and chloroplast SSRs
Authors: Salmaso Marzia , Valle Renzo Dalla , Lucchin Margherita
Published: October 2008

A germplasm safeguard programme was set up with 19 grapevine varieties considered as indigenous to northeastern Italy. To better estimate how genetic structure can be used to obtain a conservation perspective of local varieties, genetic variability was examined at 30 nuclear and 3 chloroplast polymorphic microsatellite loci in the native varieties plus 7 European cultivars taken as reference. The genetic profiles of all the cultivars were searched for possible parentage relationships and several suspected cases of the same variety having different names were investigated. The alleles shared at the loci suggest a parent–offspring relationship between Merlot and Cabernet Franc, ‘Gruaja’ and ‘Negrara Veronese’, and Marzemina Nera and Marzemina Bianca. Alleles at the 30 nuclear loci are consistent with Raboso Veronese being the progeny of Marzemina Bianca and Raboso Piave. Chloroplast-specific haplotypes were singled out for the first time in this indigenous germplasm and should be considered typical of the region. It is hypothesized that there are many specific haplotypes for the local varieties due to a past contribution of wild grapevine to the cultivated gene pool. The majority of investigated cultivars were demonstrated to constitute an independent source of genetic variation, and therefore a possible valuable resource of genetic traits for breeders.
Interaction between Hsp70 and the SR Ca2+pump: a potential mechanism for cytoprotection in heart and skeletal muscle
Authors: Tupling A. Russell , Bombardier Eric , Vigna Chris , Quadrilatero Joe , Fu Minghua
Published: October 2008

The overexpression of heat shock protein 70 (Hsp70) provides cytoprotection to cells, making them resistant to otherwise lethal levels of stress. In this review, the role Hsp70 plays in protecting both cardiac and skeletal muscle against the pathophysiological effects of oxidative stress are examined, with a focus on the molecular basis for the cytoprotective effects of Hsp70. The ability of Hsp70 to maintain cell survival undoubtedly involves the regulation of multiple steps within apoptotic pathways, but could also involve the regulation of key upstream mediators of apoptosis (i.e., oxidative stress, Ca2+overload). Hsp70 can stabilize the structure and function of both the skeletal muscle and cardiac Ca2+pump under heat stress conditions. Given that Ca2+overload has long been implicated in cell death, Hsp70 might protect muscle cells by maintaining cellular Ca2+homeostasis, thereby preventing the initiation of apoptosis. The functional interaction between Hsp70 and Ca2+pumps might also promote improvements in muscle contractility after exposure to oxidative stress.