bluuebunny/crossref_metadata_2025 huggingface dataset:
Natural Infection of Iris yellow spot virus in Twoscale Saltbush (Atriplex micrantha) Growing in Utah
Authors: Evans C. K. , Bag S. , Frank E. , Reeve J. R. , Ransom C. , Drost D. , Pappu H. R.
Published: March 2009

Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) continues to be an economically important pathogen affecting onion bulb and seed production in several parts of the world and the United States (1). Several weeds were reported naturally infected with IYSV (1,2,4). Leaves of Atriplex micrantha Ledeb. (synonym A. heterosperma Bunge) were collected from naturally occurring plants in a weed trial conducted in commercial onions grown in Box Elder County, UT on 24 September 2008. Leaves displayed a range of symptoms including spotting, chlorosis, and necrosis. Symptomatic leaves were preferentially selected for subsequent diagnostic analyses. Samples were positive for IYSV when tested by double-antibody sandwich-ELISA using a commercially available kit (Agdia Inc., Elkhart, IN). For further confirmation, total nucleic acid extracts from the symptomatic parts of the leaves were prepared and tested for the presence of IYSV by reverse transcription-PCR with primers specific to the nucleocapsid (N) gene coded by the small (S)-RNA of IYSV. The forward and reverse primer pair, 5′-TCAGAAATCGAGAAACTT-3′ and 5′-CACCAATGTCTTCAACAATCTT-3′, respectively, amplifies a 751-nt fragment of the N gene (3). An amplicon of expected size was obtained, cloned, and sequenced. The nucleotide sequence analysis and comparison with known IYSV S-RNA sequences showed that the sequence of the amplicon from A. micrantha (GenBank Accession No. FJ493541) shared more than 84% nt sequence identity with the corresponding region of IYSV isolates available in GenBank, confirming the IYSV infection of the new host weed. The highest sequence identity (98%) was with an IYSV isolate from Jefferson County, OR (GenBank Accession No. DQ233479). To our knowledge, this is the first report of IYSV infection of A. micrantha under natural conditions. The role of A. micrantha and other weeds in IYSV epidemiology needs further investigation.

References: (1) D. Gent et al. Plant Dis. 90:1468, 2006. (2) C. Nischwitz et al. Plant Dis. 91:1518, 2007. (3) H. R. Pappu et al. Arch. Virol. 151:1015, 2006. (4) R. Sampangi et al. Plant Dis. 91:1683, 2007.
First Report of Impatiens necrotic spot virus in Blackberry in the Southeastern United States
Authors: Tzanetakis I. E. , Guzmán-Baeny T. L. , VanEsbroeck Z. P. , Fernandez G. E. , Martin R. R.
Published: March 2009

Blackberry yellow vein disease (BYVD) has emerged as an important disease of blackberry (Rubus spp.) in the south and southeastern United States (2,3). In an effort to characterize viruses that may be involved in the disease, double-stranded RNA extracted from a symptomatic ‘Apache’ blackberry from South Carolina was used for shotgun cDNA cloning (4). Sequence analysis showed that in addition to Blackberry yellow vein associated virus (BYVaV) (2), a constant component of BYVD, sequences of Impatiens necrotic spot virus (INSV) also were obtained. The 623-nt fragment of INSV (Genbank Accession No. EU287930) shared 98% nucleotide and amino acid sequence identity with GenBank Accession No. NC003616. Confirmation of the results of the initial shotgun cloning was done by reverse transcription-PCR with primers INSVF (5′ GATCTGTCCTGGGATTGTTC 3′) and INSVR (5′ GTCTCCTTCTGGTTCTATAATCAT 3′) that amplify a 460 base fragment of the M RNA of INSV. Amplicons obtained from single-stranded and dsRNA templates were sequenced and found to be identical with EU287930. The identity of INSV by PCR was also supported by positive results with a commercially available INSV-ELISA kit (AC Diagnostics, Fayetteville, AR). Earlier, more than 400 plants from North Carolina, South Carolina, and Virginia with BYVD and other virus-like symptoms were tested for INSV by ELISA and approximately 33% were found to be infected with the virus (1). Thus, INSV appears to be one of the major viruses infecting blackberry in the southeastern United States, and it remains to be seen if INSV acts synergistically with BYVaV and other viruses to contribute to the severity of BYDV. To our knowledge, this is the first report of INSV infecting Rubus spp.

References: (1) T. L. Guzmán-Baeny. M.S. thesis. North Carolina State University, Raleigh, 2003. (2) J. Susaimuthu et al. Plant Pathol. 55:607, 2006. (3) J. Susaimuthu et al. Virus Res. 131:145, 2008. (4) I. E. Tzanetakis et al. J. Virol. Methods 124:73, 2005.
First Report of Crown Rot, Stem Rot, and Root Rot Caused by Binucleate Rhizoctonia AG-G on Viburnum tinus in Italy
Authors: Polizzi G. , Aiello D. , Vitale A. , Lahoz E. , Nicoletti R. , Hyakumachi M.
Published: March 2009

Laurustinus (Viburnum tinus L.), native to the Mediterranean Region, is an evergreen shrub belonging to the Caprifoliaceae that is commonly cultured as an ornamental shrub or small tree. During the summer and autumn of 2007 and 2008, a widespread yellowing, partial foliar necrosis, or death of the whole plant was observed on 3- to 4-year-old potted plants of V. tinus in a commercial nursery in eastern Sicily (Italy). More than 20% of the plants showed disease symptoms. Infected roots, crowns, and stems turned dark brown, leaves gradually became necrotic, and infected plants were often killed. Diseased tissues were disinfested for 1 min in 1% NaOCl, rinsed in sterile water, plated on potato dextrose agar (PDA) amended with streptomycin sulfate at 100 mg/liter, and then incubated at 25°C. A binucleate Rhizoctonia (BNR) species was consistently isolated from affected tissue of Laurustinus. Fungal colonies were initially white, then turned light brown or brown with age, and formed irregularly shaped, light brown sclerotia after 10 days. Hyphal cells were determined to be binucleate when stained with 1% safranin O and 3% KOH solution (1) and examined with a microscope at ×400. Anastomosis groups were determined by pairing isolates on 2% water agar in petri plates (4). Pairings were made with tester strains of binucleate Rhizoctonia AG-A through AG-S, except AG-J and AG-M. Anastomosis was observed only with tester isolates of AG-G. The rDNA-ITS of two isolates of BNR (DISTEF-Vt 31 and DISTEF-Vt 32) was sequenced (GenBank Accession Nos. AB478783 and AB478784, respectively) (3). The sequence from these two isolates exhibited 100% homology with BNR AG-G (GenBank Accession No. AY927334). Pathogenicity tests were conducted on potted, healthy, 6-month-old laurustinus. Twenty plants were inoculated by placing 1-cm2 plugs of PDA from 5-day-old mycelial cultures near the base of the stem. The same number of plants was treated with 1-cm2 PDA plugs as control. Plants were kept at 25°C and 95% relative humidity with a 12-h fluorescent light/dark regimen. Stem, crown, and root rot symptoms, identical to ones observed in nursery, appeared 20 days after inoculation, and all the inoculated plants showed symptoms within 1 month. Control plants remained healthy. Binucleate Rhizoctonia was reisolated from symptomatic tissues, completing Koch's postulates. R. solani was previously reported on Viburnum sp. in the United States (2). To our knowledge, this is the first report of binucleate Rhizoctonia causing disease on V. tinus.

References: (1) R. J. Bandoni. Mycologia 71:873, 1979. (2) D. F. Farr et al. Page 1252 in: Fungi on Plants and Plant Products in the United States. The American Phytopathological Society. St. Paul, MN, 1989. (3) M. Hyakumachi et al. Phytopathology 95:784, 2005. (4) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.
Identifying and Quantifying Phakopsora pachyrhizi Spores in Rain
Authors: Barnes C. W. , Szabo L. J. , Bowersox V. C.
Published: March 2009

In summers of 2005 and 2006, rain was collected weekly at over 100 selected National Atmospheric Deposition Program/National Trends Network sites across the soybean-growing region of the central and eastern United States. Rain samples were screened for Phakopsora pachyrhizi (causal agent of soybean rust) DNA using a nested real-time polymerase chain reaction assay. Over this time frame, P. pachyrhizi spores were detected in every state in the study, but more frequently in states along the Gulf and Atlantic coasts and along the Ohio River Valley westward to Kansas. A bimodal temporal distribution of samples testing positive for P. pachyrhizi was found in both years. However, there was a greater than threefold increase in the number of samples testing positive for P. pachyrhizi in 2006 compared with 2005, with the most significant increase in August. There was also an increase in the average number of spores per sample in 2006 relative to 2005. Sequence analysis of a subset of positive samples was used to validate the assay results. From the sequence analysis, two reliable polymorphic regions were found, resulting in six distinct genotypes. One genotype was found in 56% of the samples tested, whereas the other genotypes were found less frequently.
Detection, Distribution, and Genetic Variability of European mountain ash ringspot-associated virus
Authors: Kallinen A. K. , Lindberg I. L. , Tugume A. K. , Valkonen J. P. T.
Published: March 2009

European mountain ash ringspot-associated virus (EMARAV) was recently characterized from mountain ash (rowan) (Sorbus aucuparia) in Germany. The virus belongs tentatively to family Bunyaviridae but is not closely related to any classified virus. How commonly EMARAV occurs in ringspot disease (EMARSD) affected mountain ash trees was not reported and was investigated here. Virus-specific detection tools such as reverse transcription-polymerase chain reaction and dot blot hybridization using digoxigenin-labeled RNA probes were developed to test 73 mountain ash trees including 16 trees with no virus-like symptoms from 16 districts in Finland and Viipuri, Russia. All trees were infected with EMARAV. Hence, EMARAV is associated with EMARSD and can also cause latent infections in mountain ash. Symptom expression and the variable relative concentrations of viral RNA detected in leaves showed no correlation. Infectious EMARAV was detected also in dormant branches of trees in winter. Subsequently, genetic variability, geographical differentiation, and evolutionary selection pressures were investigated by analyzing RNA3 sequences from 17 isolates. The putative nucleocapsid (NP) gene sequence (944 nucleotides) showed little variability (identities 97 to 99%) and was under strong purifying selection. Amino acid substitutions were detected in two positions at the N terminus and one position at the C terminus of NP in four isolates. The 3′ untranslated region (442 nucleotides) was more variable (identities 94 to 99%). Six isolates from a single sampling site exhibited as wide a genetic variability as isolates from sites that were hundreds of kilometers apart and no spatial differentiation of populations of EMARAV was observed.
Pathogenic Variation ofPhakopsora pachyrhiziInfecting Soybean in Nigeria
Authors: Twizeyimana M. , Ojiambo P. S. , Sonder K. , Ikotun T. , Hartman G. L. , Bandyopadhyay R.
Published: March 2009

Soybean rust, caused by Phakopsora pachyrhizi, is an important disease in Nigeria and many other soybean-producing countries worldwide. To determine the geographical distribution of soybean rust in Nigeria, soybean fields were surveyed in the Derived Savanna (DS), Northern Guinea Savanna (NGS), and Southern Guinea Savanna (SGS) agroecological zones in Nigeria between 2004 and 2006. Disease severity in each zone was determined and analyzed using geostatistics. Prevalence of infected fields and disease severity in surveyed fields were significantly (P < 0.05) different between geographical zones with both variables being higher in the DS zone than in either NGS or SGS zones. Geostatistical analysis indicated that the spatial influence of disease severity at one location on severity at other locations was between 75 and 120 km. An exponential model best described the relationship between semivariance and lag distance when rust severity was high. Spatial interpolation of rust severity showed that locations in the DS zone were more conducive for the rust epidemic compared to areas in the NGS zone. In the 2005 survey, 116 purified isolates were established in culture on detached soybean leaves. To establish the nature of pathogenic variation in P. pachyrhizi, a set of four soybean accessions with Rpp1, Rpp2, Rpp3, and Rpp4resistance genes, two highly resistant and two highly susceptible genotypes were inoculated with single uredinial isolates. Principal component analysis on the number of uredinia per square centimeter of leaf tissue for 116 isolates indicated that an adequate summary of pathogenic variation was obtained using only four genotypes. Of these four, PI 459025B (with Rpp4gene) and TG× 1485-1D had the lowest and highest number of uredinia per square centimeter, respectively. Based on cluster analysis of the number of uredinia per square centimeter, seven pathotype clusters were determined. Isolates in cluster III were the most virulent, while those in cluster IV were the least virulent. Shannon's index (H) revealed a more diverse pathogen population in the DS zone (H = 1.21) compared to the rust population in SGS and NGS with H values of 1.08 and 0.91, respectively. This work will be useful in breeding and management of soybean rust by facilitating identification of resistant genotypes and targeting cultivars with specific resistance to match prevailing P. pachyrhizi pathotypes in a given geographical zone.
Function of Genes Encoding Acyl-CoA Synthetase and Enoyl-CoA Hydratase for Host-Selective ACT-Toxin Biosynthesis in the Tangerine Pathotype of Alternaria alternata
Authors: Miyamoto Y. , Ishii Y. , Honda A. , Masunaka A. , Tsuge T. , Yamamoto M. , Ohtani K. , Fukumoto T. , Gomi K. , Peever T. L. , Akimitsu K.
Published: March 2009

The tangerine pathotype of Alternaria alternata produces host-selective ACT-toxin and causes Alternaria brown spot disease. Sequence analysis of a genomic cosmid clone identified a part of the ACTT gene cluster and implicated two genes, ACTT5 encoding an acyl-CoA synthetase and ACTT6 encoding an enoyl-CoA hydratase, in the biosynthesis of ACT-toxin. Genomic Southern blots demonstrated that both genes were present in tangerine pathotype isolates producing ACT-toxin and also in Japanese pear pathotype isolates producing AK-toxin and strawberry pathotype isolates producing AF-toxin. ACT-, AK-, and AF-toxins from these three pathotypes share a common 9,10-epoxy-8-hydroxy-9-methyl-decatrienoic acid moiety. Targeted gene disruption of two copies of ACTT5 significantly reduced ACT-toxin production and virulence. Targeted gene disruption of two copies of ACTT6 led to complete loss of ACT-toxin production and pathogenicity and a putative decatrienoic acid intermediate in ACT-toxin biosynthesis accumulated in mycelial mats. These results indicate that ACTT5 and ACTT6 are essential genes in ACT-toxin biosynthesis in the tangerine pathotype of A. alternata and both are required for full virulence of this fungus.
Modeling Disease Progression of Camellia Twig Blight Using a Recurrent Event Model
Authors: Thomson J. L. , Copes W. E.
Published: March 2009

To improve control of camellia twig blight (CTB) using sanitation methods, a more complete epidemiologic understanding of this disease is necessary. Three CTB disease stages were modeled using recurrent event analysis. Wound inoculated stems were observed at regular intervals for appearance of disease symptoms. Survival times (time from inoculation until symptom appearance) for the three disease stages (mild, moderate, and severe) were regressed against stem diameter, monthly mean hours/day within a specified temperature range (15 to 30°C), and season (spring, summer, fall, and winter). For all three CTB disease stages, stem diameter had a protective effect on survival times, while monthly mean hours/day in the specified temperature range and warmer seasons were risk factors. Based upon median ratios, the mild disease stage developed 2 to 3 times faster in spring, summer, and fall than in winter. Similarly, moderate and severe disease stages developed 2 to 2.5 times faster. For all three disease stages, seasonal differences in stage development were smaller among fall, spring, and summer, varying from 1 to 1.6 times faster. Recurrent event modeling of CTB progression provides knowledge concerning developmental expression of this disease, information necessary for creating a comprehensive, integrated disease management program.
Multilocus Phylogenetic Diversity of Fusarium avenaceum Pathogenic on Lisianthus
Authors: Nalim F. A. , Elmer W. H. , McGovern R. J. , Geiser D. M.
Published: March 2009

Fusarium avenaceum is a globally distributed fungus commonly isolated from soil and a wide range of plants. Severe outbreaks of crown and stem rot of the flowering ornamental, lisianthus (Eustoma grandiflorum), have been attributed to F. avenaceum. We sequenced portions of the translation elongation factor 1-alpha (tef) and beta-tubulin (benA) protein coding genes as well as partial intergenic spacer (IGS) regions of the nuclear ribosomal genes in 37 Fusarium isolates obtained from lisianthus and other host plants. Isolates that were previously identified morphologically as F. acuminatum were included as an outgroup. Phylogenetic analyses of tef, benA, and IGS sequences showed that F. avenaceum isolates were an exclusive group with strong bootstrap support and no significant incongruence among gene genealogies. Isolates from lisianthus were scattered within this clade and did not form distinct groups based on host species or locality. Pathogenicity tests of F. avenaceum isolates obtained from several other hosts showed an ability to cause disease on lisianthus, suggesting that F. avenaceum may be pathogenic on lisianthus regardless of its phylogenetic origin. These findings have management implications and suggest that any host that supports F. avenaceum may serve as a source of inoculum for lisianthus growers.
Enhanced immune response and protection efficacy of a DNA vaccine constructed by linkage of the Mycobacterium tuberculosis Ag85B-encoding gene with the BVP22-encoding gene
Authors: Yao Wanhong , Liu Shengwu , Qu Xueju , Xiao Shaobo , Liu Yan , Liu Junyan
Published: March 2009

Plasmid DNA vaccines have been widely explored for use in tuberculosis immunization but their immunogenicity needs improvement. In the present study, we incorporated the bovine herpesvirus 1 VP22 (BVP22)-encoding gene, which encodes a protein that demonstrates a capability for disseminating the expressed antigen to neighbouring cells, into a DNA vector in which it was fused to the Ag85B-encoding gene ofMycobacterium tuberculosis(Mtb), and investigated whether this linkage could enhance immune response and protective efficacy in C57BL/6 mice compared to plasmid DNA encoding Ag85B alone. After immunization in mice, Ag85B-specific ELISA antibodies and spleen lymphocyte proliferative responses induced by DNA co-expressing BVP22 and Ag85B were significantly higher than those obtained in mice immunized with Ag85B-encoding DNA alone, except for the number of gamma interferon secreting cells. In addition, based on histopathological examination and bacterial-load determination in lung and spleen, protection against intravenous Mtb H37Rv challenge evoked by the BVP22–Ag85B DNA immunization exceeded the response elicited by Ag85B DNA alone, which was not significantly different from that provided by Bacillus Calmette–Guérin (BCG). These results suggested that DNA vaccine consisting of BVP22 and Ag85B-encoding DNA enhanced immune response and protection against intravenous Mtb H37Rv challenge in mice, indicating that BVP22-encoding DNA might be a promising tool to enhance TB DNA vaccine efficacy.
Mediterranean spotted fever with encephalitis
Authors: Aliaga Luis , Sánchez-Blázquez Patricia , Rodríguez-Granger Javier , Sampedro Antonio , Orozco Miguel , Pastor Jorge
Published: March 2009

Rickettsia conorii infection is endemic in the Mediterranean basin, where it is known as Mediterranean spotted fever, also known as Boutonneuse fever and Marseilles fever. We report the case of a 66-year-old diabetic man who presented a severe form of the disease, complicated by acute renal failure, thrombocytopenia and encephalitis. Diagnosis was confirmed by indirect immunofluorescence assay. Despite appropriate treatment, severe neurological sequelae have remained. Medical literature on encephalitis caused by R. conorii is also reviewed.
Neutrophil enhancement of Pseudomonas aeruginosa biofilm development: human F-actin and DNA as targets for therapy
Authors: Parks Quinn M. , Young Robert L. , Poch Katie R. , Malcolm Kenneth C. , Vasil Michael L. , Nick Jerry A.
Published: March 2009

In the cystic fibrosis (CF) airway, chronic infection byPseudomonas aeruginosaresults from biofilm formation in a neutrophil-rich environment. We tested the capacity of human neutrophils to modify early biofilm formation ofP. aeruginosastrain PAO1, and an isogenic CF strain isolated early and years later in infection. In a static reactor,P. aeruginosabiofilm density of all strains was enhanced at 24 h in the presence of neutrophils, with the greatest relative increase associated with the lowest inoculum ofP. aeruginosatested. Previously, neutrophil-induced biofilm enhancement was shown to largely result from the incorporation of F-actin and DNA polymers into the bacterial biofilm. This finding was advanced by the comparison of biofilm enhancement from intact unstimulated neutrophils and from lysed or apoptotic neutrophils. Apoptotic neutrophils, with an intact cell membrane, were unable to contribute to biofilm enhancement, while lysed neutrophils evoked a similar response to that of intact cells. Using F-actin and DNA as targets, the capacity of negatively charged poly(amino acids) to disrupt, or prevent, early biofilm formation was tested. Anionic poly(aspartic acid) effectively prevented or disrupted biofilm formation. Combination of poly(aspartic acid) with DNase resulted in a synergistic increase in biofilm disruption. These results demonstrate that the presence of dying neutrophils can facilitate the initial stages of biofilm development by low inocula ofP. aeruginosa. Neutrophil F-actin represents a potential new therapeutic target for disruption of pathogenic biofilms.
Prospective study of potential sources of Streptococcus mutans transmission in nursery school children
Authors: Alves Alessandra C. , Nogueira Ruchele D. , Stipp Rafael N. , Pampolini Flávia , Moraes Antonio B. A. , Gonçalves Reginaldo B. , Höfling José F. , Li Yihong , Mattos-Graner Renata O.
Published: March 2009

Transmission of Streptococcus mutans, a major dental caries pathogen, occurs mainly during the first 2.5 years of age. Children appear to acquire S. mutans mostly from their mothers, but few studies have investigated non-familial sources of S. mutans transmission. This study prospectively analysed initial S. mutans oral colonization in 119 children from nursery schools during a 1.5-year period and tracked the transmission from child to child, day-care caregiver to child and mother to child. Children were examined at baseline, when they were 5–13 months of age, and at 6-month intervals for determination of oral levels of S. mutans and development of caries lesions. Levels of S. mutans were also determined in caregivers and mothers. A total of 1392 S. mutans isolates (obtained from children, caregivers and mothers) were genotyped by arbitrarily primed PCR and chromosomal RFLP. Overall, 40.3 % of children were detectably colonized during the study, and levels of S. mutans were significantly associated with the development of caries lesions. Identical S. mutans genotypes were found in four nursery cohorts. No familial relationship existed in three of these cohorts, indicating horizontal transmission. Despite high oral levels of S. mutans identified in most of the caregivers, none of their genotypes matched those identified in the respective children. Only 50 % of children with high levels of S. mutans carried genotypes identified in their mothers. The results support previous evidence indicating that non-familial sources of S. mutans transmission exist, and indicate that this bacterium may be transmitted horizontally between children during the initial phases of S. mutans colonization in nursery environments.
The prevalence and virulence characteristics of enteroaggregative Escherichia coli at an urgent-care clinic in the USA: a case–control study
Authors: Cennimo David , Abbas Atif , Huang David B. , Chiang Tom
Published: March 2009

This case–control study examined the prevalence of enteroaggregative Escherichia coli (EAEC), its genes and elicited inflammatory response, and the stool characteristics of adult patients with and without acute diarrhoeal illness presenting to an urgent-care clinic in the USA. A total of 1004 individual stool specimens (253 from patients with acute diarrhoeal illness and 751 from patients without diarrhoeal illness) were collected between 1 June 2003 and 30 June 2008. EAEC was identified as the sole cause of acute diarrhoeal illness in 6 % (n=15) of patients and in 2 % (n=15) without diarrhoeal illness. Control patients (n=15) were similar to case patients (n=15) for age, gender and co-morbidities. The EAEC genes aggR, aap, aat, astA and/or set1A were identified more frequently in case patients compared with control patients (P <0.05). aggR-positive EAEC elicited higher levels of interleukin (IL)-1ra, IL-6, IL-8 and tumour necrosis factor-α compared with aggR-negative EAEC during co-incubation with HCT-8 cells. Patients with EAEC diarrhoea and isolates with the genes aggR, aap, aatA, astA or set1A had stools characterized by gross mucus and the presence of faecal leukocytes (P <0.05). These results indicate that EAEC is a potential cause of acute diarrhoeal illness affecting patients presenting to an acute-care clinic in the USA and suggest that aggR, aap, aatA, astA and set1A may be markers for virulence.
Aetiology of influenza-like illness in adults includes parainfluenzavirus type 4
Authors: Hasman Hatice , Pachucki Constance T. , Unal Arife , Nguyen Diep , Devlin Troy , Peeples Mark E. , Kwilas Steven A.
Published: March 2009

Influenza viruses cause significant morbidity and mortality in adults each winter. At the same time, other respiratory viruses circulate and cause respiratory illness with influenza-like symptoms. Human respiratory syncytial virus (HRSV), human parainfluenza viruses (HPIV) and human metapneumovirus have all been associated with morbidity and mortality in adults, including nosocomial infections. This study evaluated 154 respiratory specimens collected from adults with influenza-like/acute respiratory illness (ILI) seen at the Edward Hines Jr VA Hospital, Hines, IL, USA, during two successive winters, 1998–1999 and 1999–2000. The samples were tested for ten viruses in two nested multiplex RT-PCRs. One to three respiratory viruses were detected in 68 % of the samples. As expected, influenza A virus (FLU-A) infections were most common (50 % of the samples), followed by HRSV-A (16 %). Surprisingly, HPIV-4 infections (5.8 %) were the third most prevalent. Mixed infections were also relatively common (11 %). When present, HPIV infections were approximately three times more likely to be included in a mixed infection than FLU-A or HRSV. Mixed infections and HPIV-4 are likely to be missed using rapid diagnostic tests. This study confirms that ILI in adults and the elderly can be caused by HRSV and HPIVs, including HPIV-4, which co-circulate with FLU-A.
Molecular typing and virulence of enteroaggregative Escherichia coli strains isolated from children with and without diarrhoea in Rio de Janeiro city, Brazil
Authors: Regua-Mangia Adriana H. , Gomes Tânia A. T. , Vieira Mônica A. M. , Irino Kinue , Teixeira Lúcia M.
Published: March 2009

Enteroaggregative Escherichia coli (EAEC) strains have been implicated as emerging aetiological agents of diarrhoea worldwide. In the present study, 43 EAEC strains were serotyped and characterized according to random amplification of polymorphic DNA profiles, PFGE, multilocus enzyme electrophoresis (MLEE) and the presence of putative virulence genes (hly, aero, kps, fim, aggA, aafA, aggR, astA, she, aap, shf and pet). The EAEC strains consisted of a diversity of serotypes including eight O-non-typable and 35 O-typable strains arranged into 21 O : H combinations. Amplification of specific genes revealed that all strains carried at least two of the virulence sequences investigated. fim, aggR and aap were the most frequent genes in both groups studied. hly, aero and aggA sequences were more prevalent in the diarrhoeal group. kps occurred exclusively in strains isolated from symptomatic children and showed strong association with diarrhoeal disease. The molecular approaches used to investigate the relatedness among EAEC strains revealed a high degree of polymorphism, suggesting that these micro-organisms have a non-clonal origin. A closer relationship was observed among EAEC strains sharing O : H types. No significant clustering could be identified related to the virulence traits investigated; however, the she locus showed clonal distribution by MLEE typing. These results are in accordance with previous findings in revealing the conservation of particular EAEC factors, despite the high degree of diversity related to both genotypic and phenotypic markers.
Molecular and clinical analyses of the gene encoding the collagen-binding adhesin of Streptococcus mutans
Authors: Nomura Ryota , Nakano Kazuhiko , Taniguchi Naho , Lapirattanakul Jinthana , Nemoto Hirotoshi , Grönroos Lisa , Alaluusua Satu , Ooshima Takashi
Published: March 2009

Streptococcus mutans is a known pathogen of dental caries and its major cell surface antigens have been widely investigated. Recently, an approximately 120 kDa Cnm protein with binding properties to type I collagen was identified, and its encoding gene (cnm) cloned and sequenced. In the present study, we sequenced cnm from 47 different clinical S. mutans strains and found that the nucleotide alignment of the collagen-binding domain was well conserved. We devised a PCR method for identifying the cnm gene, examined the prevalence of cnm-positive S. mutans strains in various mother–child groups, and assessed the significance of such strains for transmission and dental caries. The detection rate of cnm-positive strains was significantly lower in strains isolated from Japanese children in the 2000s (8.0 %) as compared to those isolated in the 1980s (15.8 %) (P<0.05). Furthermore, the presence of S. mutans possessing cnm in salivary specimens collected from 55 S. mutans-positive mother–child pairs was 40 and 32.7 % in the mothers and children, respectively. The frequency of cnm-positive children whose mothers were also positive was 72 %, which was significantly higher than that of cnm-positive children with negative mothers (P<0.0001, odds ratio 17.5). In addition, clinical parameters indicating dental caries were significantly increased in children with cnm-positive S. mutans in saliva (n=13), as compared to those with cnm-negative S. mutans (n=15) and S. mutans-negative children (n=20) (P<0.01). These results indicate that cnm-positive S. mutans strains are closely correlated with dental caries, while vertical transmission in cnm-positive mother–child pairs was also demonstrated.
Conspicuous behaviour of Fundulus heteroclitus associated with high digenean metacercariae gill abundances
Authors: Santiago Bass C. , Weis J. S.
Published: March 2009

Fundulus heteroclitus from six sites throughout the Hackensack Meadowlands District in northern NJ, U.S.A., were examined. Differences in behaviour (surfacing, conspicuousness and activity) were compared to gill infection intensity. Fish from populations infected with >1500 digenean metacercariae of Ascocotyle phagicola diminuta and Echinochasmus schwartzi, spent significantly more time at the water surface and exhibited significantly more conspicuous behaviour (e.g. jerking) than fish from less parasitized populations. This more conspicuous behaviour has the potential to increase trophic transmission of the parasite to its definitive wading‐bird host.
The effect of nest aggregation on the reproductive behaviour of the peacock blenny Salaria pavo
Authors: Saraiva J. L. , Barata E. N. , Canário A. V. M. , Oliveira R. F.
Published: March 2009

The effect of nest aggregation in courtship behaviour was tested experimentally in an ecologically constrained, sex‐role reversed population of the peacock blenny Salaria pavo. Mixed sex groups of eight males and eight females were tested in experimental tanks, containing eight potential nests either aggregated or dispersed. In the aggregated treatment, males spent more time inside their nests and monopolized other potential nests, causing a female‐biased operational sex ratio (OSR). In the aggregated treatment, females also expressed more courtship behaviour. The results in general support the prediction that the aggregation of nests promotes male monopolization of potential nests, resulting in fewer nest‐holding males and therefore a female‐biased OSR that leads to the reversal of sex roles.
Establishment of dominance relationships in gilthead sea bream Sparus aurata juveniles during feeding: effects on feeding behaviour, feed utilization and fish health
Authors: Montero D. , Lalumera G. , Izquierdo M. S. , Caballero M. J. , Saroglia M. , Tort L.
Published: March 2009

In gilthead sea bream Sparus aurata held in groups of two, five or 10 fish, social hierarchies were observed. Subordinate S. aurata were characterized by elevation of basal levels of plasma cortisol, together with a reduced immunological potential. Subordinate fish also showed lower feed intake, feed utilization and lower growth. Fatty acid composition was also affected by social status, with a lower content of saturated acids, oleic and eicosapentaenoic, in muscle and liver of fish considered as subordinate. Results show that social hierarchy acts as a stressor in S.aurata.
Seasonal changes in antioxidant defence system of liver and gills of Salmo trutta caspius, Salmo trutta labrax and Salmo trutta macrostigma
Authors: Aras N. M. , Bayir A. , Sirkecioglu A. N. , Bayir M. , Aksakal E. , Haliloglu H. I.
Published: March 2009

Seasonal changes in antioxidant enzyme activities (superoxide dismutase, SOD, EC 1.15.1.1; catalase, CAT, EC 1.11.1.16; glutathione peroxidase, GPx, EC 1.11.1.9; glutathione reductase, GR, EC 1.6.4.2; glucose‐6‐phosphate dehydrogenase, G6PD, EC 1.1.1.49 and glutathione S‐transferase, GST, EC 1.5.1.18) and lipid peroxidation (LPO) levels of livers and gills of female Caspian trout Salmo trutta caspius, Black Sea trout Salmo trutta labrax and mountain trout Salmo trutta macrostigma were investigated. SOD, CAT, GPx, G6PD and GST activities were higher in liver compared to gills of all sub‐species; concomitantly, the GR activity was also higher in the livers of S. t. caspius and S. t. labrax, but the reverse was seen in S. t. macrostigma. LPO levels were higher in the gills compared to the liver of all sub‐species. There was no general trend in the seasonal changes in the gill antioxidant enzyme (AE) activities or LPO levels. Higher AE activities, however, were found in the liver of each sub‐species during autumn, and this coincided with an increase in the gonado‐somatic index.
Depth distribution and biological characteristics of the European eel Anguilla anguilla in Lough Ennell, Ireland
Authors: Yokouchi K. , Aoyama J. , Miller M. J. , McCarthy T. K. , Tsukamoto K.
Published: March 2009

Using a longline survey, a total of 196 European eels Anguilla anguilla were collected at different depths in Lough Ennell (maximum depth 30 m), central Ireland. The catch per unit of effort of A. anguilla that were caught from 1 to 25 m depths was lowest at 0·5–5·0 m and greatest at the deepest depth range (22·5–25·0 m). Sub‐samples of A. anguilla from depths of <15 m showed little or no difference in size, sex ratio, age, growth rate, condition factor, length–mass relationship, gonado‐somatic index, fin index or eye index with fish from depths of >15 m. All fish examined were female yellow‐phase A. anguilla that had ages from 7 to 20 years (mean ±s.d. = 10·3 ± 2·9 years), with growth rates from 24·0–60·8 mm year−1 (mean ±s.d. = 40·7 ± 8·5 mm year−1). Variations in the growth rates were greater in the shallow group than that of the deep group. This study suggested that deeper regions are important feeding habitats for A. anguilla and that fish in this lake were growing moderately fast compared to similar habitats and areas in the species’ range.
A comparison of gut evacuation models for larval mackerel (Scomber scombrus) using serial photography
Authors: Dunbrack R. , Giguère L. A. , St‐Pierre J.‐F.
Published: March 2009

A novel technique is described, using serial photography of the gut contents of transparent living larval fishes, to generate individual gut evacuation time series. This technique was applied to Atlantic mackerel Scomber scombrus larvae to compare three widely used models of gut evacuation: linear, exponential and square‐root. Regression r2 for the exponential model exceeded those for the linear and square root models in 20 of 21 time series, strongly supporting the exponential model. At the initial gut fullness for each time series, total gut evacuation rates calculated with the exponential model averaged 2·2 and 1·3 times greater than those calculated with the linear and square‐root models, respectively, and would produce correspondingly higher estimates of feeding rates for field‐collected larvae with similar levels of gut fullness. The results highlight the importance of choosing the appropriate evacuation model in feeding studies, particularly those intended to examine short‐term changes in larval fish feeding rates, a contributing factor to the highly variable yearly recruitment of many marine fish species.
Pathogenetic significance of ecotropic viral integration site‐1 in hematological malignancies
Authors: Goyama Susumu , Kurokawa Mineo
Published: March 2009

The ecotropic viral integration site‐1 (Evi‐1) gene was first identified as a common locus of retroviral integration in murine leukemia models. In humans, EVI‐1 is located on chromosome 3q26, and rearrangements on chromosome 3q26 often activate EVI‐1 expression in hematological malignancies. Overexpression of EVI‐1 also occurs with high frequency in leukemia patients without 3q26 abnormalities, and importantly, high EVI‐1 expression is an independent negative prognostic indicator irrespective of the presence of 3q26 rearrangements. Recent gene targeting studies in mice revealed that Evi‐1 is preferentially expressed in hematopoietic stem cells and plays an essential role in proliferation and maintenance of hematopoietic stem cells. In addition, intense attention has been focused on the EVI‐1 gene complex as retrovirus integration sites because transcription‐activating integrations into the EVI‐1 locus confer survival and self‐renewing ability to hematopoietic cells. The experimental results using animal models suggest that activation of Evi‐1 in hematopoietic cells leads to clonal expansion or dysplastic hematopoiesis, whereas onset of full‐blown leukemia requires cooperative genetic events. EVI‐1 possesses diverse functions as an oncoprotein, including suppression of transforming growth factor‐β‐mediated growth inhibition, upregulation of GATA2, inhibition of the Jun kinase pathway, and stimulation of cell growth via activator protein‐1. In this article, we summarize current knowledge regarding the biochemical properties and biological functions of EVI‐1 in normal and malignant hematopoiesis, with specific focus on its pathogenetic significance in hematological malignancies. (Cancer Sci 2009; 100: 990–995)
Assignment of 128 genes localized on human chromosome 14q to the IMpRH map
Authors: Shimogiri T. , Nishibori M. , Hiraiwa H. , Hayashi T. , Yasue H.
Published: March 2009

To provide a gene‐based comparative map and to examine a porcine genome assembly using bacterial artificial chromosome‐based sequence, we have attempted to assign 128 genes localized on human chromosome 14q (HSA14q) to a porcine 7000‐rad radiation hybrid (IMpRH) map. This study, together with earlier studies, has demonstrated the following. (i) 126 genes were incorporated into two SSC7 RH linkage groups by CarthaGene analysis. (ii) In the remaining two genes, TOX4 linked to TCRA located in SSC7 by two‐point analysis, whereas SIP1 showed no significant linkage with any gene/marker registered in the IMpRH Web Server. (iii) In the two groups, the gene clusters located from 19.9 to 36.5 Mb on HSA14q11.2‐q13.3 and from 64.0 to 104.3 Mb on HSA14q23‐q32.33 respectively were assigned to SSC7q21‐q26. (iv) Comparison of the gene order between the present RH map and the latest porcine sequence assembly revealed some inconsistencies, and a redundant arrangement of 16 genes in the sequence assembly.
The impact of dose escalation and resistance modulation in older patients with acute myeloid leukaemia and high risk myelodysplastic syndrome: the results of the LRF AML14 trial
Authors: Burnett Alan K. , Milligan Donald , Goldstone Anthony , Prentice Archibald , McMullin Mary‐Frances , Dennis Michael , Sellwood Elizabeth , Pallis Monica , Russell Nigel , Hills Robert K. , Wheatley Keith ,   on behalf of the United Kingdom National Cancer Research Institute Haematological Oncology Study Group
Published: March 2009

The acute myeloid leukaemia (AML)14 trial addressed four therapeutic questions in patients predominantly aged over 60 years with AML and High Risk Myelodysplastic Syndrome: (i) Daunorubicin 50 mg/m2 vs. 35 mg/m2; (ii) Cytarabine 200 mg/m2 vs. 400 mg/m2 in two courses of DA induction; (iii) for part of the trial, patients allocated Daunorubicin 35 mg/m2 were also randomized to receive, or not, the multidrug resistance modulator PSC‐833 in a 1:1:1 randomization; and (iv) a total of three versus four courses of treatment. A total of 1273 patients were recruited. The response rate was 62% (complete remission 54%, complete remission without platelet/neutrophil recovery 8%); 5‐year survival was 12%. No benefits were observed in either dose escalation randomization, or from a fourth course of treatment. There was a trend for inferior response in the PSC‐833 arm due to deaths in induction. Multivariable analysis identified cytogenetics, presenting white blood count, age and secondary disease as the main predictors of outcome. Although patients with high Pgp expression and function had worse response and survival, this was not an independent prognostic factor, and was not modified by PSC‐833. In conclusion, these four interventions have not improved outcomes in older patients. New agents need to be explored and novel trial designs are required to maximise prospects of achieving timely progress.
The evaluation of human papillomavirus genotyping in cervical liquid‐based cytology specimens; using the Roche Linear Array HPV genotyping assay
Authors: Jamison J. , Wilson R. T. , Carson J.
Published: March 2009

Objective:  To ascertain the usefulness of the Roche Linear Array human papillomavirus (HPV) genotyping assay for assessing HPV genotypes in liquid‐based cytology (LBC) samples and to evaluate this methodology within a cytopathology laboratory. These tests are of importance as persistent infection with high‐risk HPV genotypes is considered a causal factor in the development of cervical cancer.

Methods:  A total of 175 cervical LBC samples were tested using the Roche Linear Array HPV genotyping test. The suitability of the assay use in routine cytopathology laboratory was considered. HPV genotypes were matched to the cervical cytology results, which included negative, borderline nuclear abnormalities, mild, moderate and severe dyskaryosis.

Results:  The assay could be applied to screening samples with the combined result available at the reporting stage. There were no test failures. All samples used after cytological analysis had sufficient DNA for testing. The results were reproducible and easily read and there was concordance of results between biomedical scientists. The results of the assay showed co‐infection with multiple HPV genotypes was common in both high‐grade and low‐grade cytology samples. The percentage of HPV+ samples in the normal cytology samples (although in this grouping the number of samples was low) was 37%. In the cytology samples reported as severe dyskaryosis the HPV genotypes most commonly found were HPV16 and HPV51.

Conclusion:  The assay was able to detect multiple HPV infection with a wide range of genotypes in LBC samples sent for routine cytological analysis. It would be suitable for use in a cytopathology laboratory. The results of the assay show that the genotype profile has some variation from other geographical regions, and more work is needed to determine population prevalence, to ascertain the impact of the HPV vaccine, to evaluate test for cure and HPV triage management.
Significance of collagenous and mucinous spherulosis in breast cytology specimens
Authors: Hata S. , Kanomata N. , Kozuka Y. , Fukuya M. , Ohno E. , Moriya T.
Published: March 2009

S. Hata, N. Kanomata, Y. Kozuka, M. Fukuya, E. Ohno and T. Moriya Significance of collagenous and mucinous spherulosis in breast cytology specimens

Objective:  Spherulosis of the breast is a rare but distinct benign morphological entity. As there are few cytological reports of breast spherulosis, the significance of spherulosis among cytological specimens is unclear. The objective was to document cytological aspects of spherulosis.

Methods:  A total of 3491 consecutive breast fine needle aspiration cytology (FNAC) samples and 69 nipple discharge cytology samples were reviewed. Papanicolaou‐stained slides with or without Romanowsky staining were analysed. The corresponding 1926 histological specimens were also reviewed.

Results:  We detected 17 cases of collagenous spherulosis (CS) and/or mucinous spherulosis (MS) among 3560 breast cytology specimens (0.48%). All samples were from women, who varied in age from 22 to 69 years. CS and/or MS were present in 15 of 3491 FNAC specimens (0.43%) and in two of 69 nipple discharge cytology specimens (2.9%). Corresponding histological specimens were available for 14 of the 17 specimens. Of the 14 specimens, 12 consisted of intraductal papilloma, one of fibroadenoma, and one of fibrocystic change. There was no spherulosis among the 1251 cytological specimens of malignant diseases.

Conclusions:  Cytological evidence of spherulosis is a good indicator of intraductal papilloma.