bluuebunny/crossref_metadata_2025 huggingface dataset:
Minimal residual disease in chronic lymphocytic leukaemia: is it ready for primetime?
Authors: Nabhan Chadi , Coutré Steven , Hillmen Peter
Published: November 2006

New therapeutic modalities have substantially improved response rates and outcomes in chronic lymphocytic leukaemia (CLL), yet the mindset remains that palliation is the only goal of therapy because the disease is considered incurable. Ultimately, all patients relapse despite achieving an initial response, as minimal residual disease (MRD) persisting after therapy eventually evolves into morphological and clinical recurrence. The emergence of immune‐based combination therapies capable of inducing molecular remissions, the availability of highly sensitive assays that detect MRD, and emerging data showing a longer duration of response or longer survival in patients with no detectable disease, suggest that eradicating MRD may be a reasonable option for some patients. Moreover, novel biological prognostic markers have divided CLL into favourable and unfavourable subtypes, arguing in favour of defining different goals of therapy for different patients. Clinicians are increasingly challenged with the task of how best to incorporate MRD assessment into clinical practice, especially in an era when these novel prognostic factors exist. This review summarises the current understanding of MRD from a clinical standpoint, suggests that MRD eradication maybe a reasonable option for some patients, and argues in favour of designing large randomised studies to determine whether MRD‐negative remission improves outcome.
Intestinal helminth co-infection has a negative impact on both anti-Mycobacterium tuberculosis immunity and clinical response to tuberculosis therapy
Authors: Resende Co T , Hirsch C S , Toossi Z , Dietze R , Ribeiro-Rodrigues R
Published: November 2006

The impact of intestinal helminth infection on Mycobacterium tuberculosis (MTB)-specific immune responses during active tuberculosis (TB) is not known. We investigated the role of intestinal helminth infection in anti-MTB immunity by evaluating both cellular phenotype and cytokine profiles in patients with TB and patients with concomitant TB and intestinal helminth infection (TB + Helm) during TB therapy. Twenty-seven per cent of TB patients enrolled for the study were co-infected with at least one intestinal helminth. At baseline, absolute frequencies of leucocytes, monocytes and eosinophils from TB and TB + Helm patients differed from healthy subjects. Concomitant intestinal helminth infection in TB + Helm patients had a negative impact (P < 0·05) on absolute frequencies of CD3+, CD4+, CD8+, natural killer (NK) T and CD4+ CD25high T cell subsets when compared to either TB patients or healthy controls. Differences in CD4+ T cell frequencies were accompanied by lower interferon (IFN)-γ and elevated and sustained interleukin (IL)-10 levels in whole blood (WB) cultures from TB + Helm compared to TB patients. In addition to a depressed anti-MTB immunity, TB + Helm patients also presented with more severe radiological pulmonary disease, with a significant difference (P = 0·013) in the number of involved lung zones at the end of TB treatment. The above data may indicate that concomitant intestinal helminth infection in patients with newly diagnosed TB skews their cytokine profile toward a T helper 2 response, which could favour persistent MTB infection and a more protracted clinical course of the disease.
Protein array autoantibody profiles for insights into systemic lupus erythematosus and incomplete lupus syndromes
Authors: Li Q-Z , Zhou J , Wandstrat A E , Carr-Johnson F , Branch V , Karp D R , Mohan C , Wakeland E K , Olsen N J
Published: November 2006

The objective of this study was to investigate the prevalence and clinical significance of a spectrum of autoantibodies in systemic lupus erythematosus and incomplete lupus syndromes using a proteome microarray bearing 70 autoantigens. Microarrays containing candidate autoantigens or control proteins were printed on 16-section slides. These arrays were used to profile 93 serum samples from patients with systemic lupus erythematosus (SLE (n = 33), incomplete LE (ILE; n = 23), first-degree relatives (FDRs) of SLE patients (n = 20) and non-autoimmune controls (NC; n = 17). Data were analysed using the significance analysis of microarray (SAM) and clustering algorithms. Correlations with disease features were determined. Serum from ILE and SLE patients contained high levels of IgG autoantibodies to 50 autoantigens and IgM autoantibodies to 12 autoantigens. Elevated levels of at least one IgG autoantibody were detected in 26% of SLE and 19% of ILE samples; elevated IgM autoantibodies were present in 13% of SLE and 17% of ILE samples. IgG autoantibodies segregated into seven clusters including two specific for DNA and RNA autoantigens that were correlated with the number of lupus criteria. Three IgG autoantibody clusters specific for collagens, DNA and histones, were correlated with renal involvement. Of the four IgM autoantibody clusters, two were correlated negatively with the number of lupus criteria; none were correlated with renal disease. The IgG : IgM autoantibody ratios generally showed a stepwise increase in the groups following disease burden from NC to SLE. Insights derived from the expanded autoantibody profiling made possible with the antigen array suggest differences in autoreactivity in ILE and SLE. Determining whether the IgM aurotreactivity that predominates in ILE represents an early stage prior to IgG switching or is persistent and relatively protective will require further longitudinal studies.
Interferon-β and adhesion molecules (E-selectin and s-intracellular adhesion molecule-1) are detected in sera from patients with retinal vasculitis and are induced in retinal vascular endothelial cells by Toll-like receptor 3 signalling
Authors: Lee M T , Hooper L C , Kump L , Hayashi K , Nussenblatt R , Hooks J J , Detrick B
Published: November 2006

Retinal vasculitis is a major component of ocular inflammation that plays a role in retinal tissue damage in patients with idiopathic uveitis and Behçet's disease. Here we show that type 1 interferons (IFN α/β) were not detected in sera from normal individuals but were identified in up to 46% of the sera from retinal vasculitis patients. The predominant form of IFN observed was IFN-β, which was detected in 39% of Behçet's disease patients and 47% of idiopathic uveitis patients. Seven patients whose sera contained IFN-β were monitored prospectively. IFN-β was shown to be present for 6–12 months in all seven of the sera samples tested. Furthermore, the adhesion molecule profile identified in this study was strikingly different when Behçet's and uveitis patient sera were compared to sera from normal controls. Sera from Behçet's disease patients contained significantly elevated levels of the soluble adhesion molecules, sE-selectin and s-intracellular adhesion molecule-1 (sICAM-1), whereas sera from patients with idiopathic uveitis contained significantly increased sE-selectin. In vitro studies evaluating the cell source of these cytokines revealed that polyriboinosinic polyribocytidylic acid (poly I:C) activated retinal vascular endothelial cells produce sE-selectin, sICAM-1 and IFN-β. Production of these molecules was inhibited by pretreatment with anti-Toll-like receptor 3 (TLR-3) antibody. In conclusion, IFN-β, sE-selectin and sICAM-1 are elevated in patients with retinal vasculitis and are induced in retinal vascular endothelial cells in vitro by activating the innate immune system through TLR-3. Further analysis of innate immune signalling may prove to be a novel target for future studies on pathogenic mechanisms and therapeutic approaches in retinal vasculitis.
The dual role of p55 tumour necrosis factor-α receptor inActinobacillus actinomycetemcomitans-induced experimental periodontitis: host protection and tissue destruction
Authors: Garlet G P , Cardoso C R B , Campanelli A P , Ferreira B R , Avila-Campos M J , Cunha F Q , Silva J S
Published: November 2006

Inflammatory immune reactions in response to periodontopathogens are thought to protect the host against infection, but may trigger periodontal destruction. Thus, we examined the mechanisms by which the proinflammatory cytokine tumour necrosis factor (TNF)-α modulates the outcome of Actinobacillus actinomycetemcomitans-induced periodontal disease in mice. Our results showed that TNF-α receptor p55-deficient mice [p55TNF-knock-out (KO)] developed a less severe periodontitis in response to A. actinomycetemcomitans infection, characterized by significantly less alveolar bone loss and inflammatory reaction. Real-time polymerase chain reaction (PCR) demonstrated that levels of chemokines (CXCL1, 3 and 10; CCL3 and 5) and their receptors (CXCR2 and 3, CCR5) were lower in p55TNF-KO mice, as were matrix metalloproteinase (MMP)-1, 2 and 9 and receptor activator of nuclear factor kB ligand (RANKL) mRNA levels. However, the absence of the TNF-α p55 results in an impairment of protective immunity to A. actinomycetemcomitans infection, characterized by increased bacterial load and higher levels of C-reactive protein during the course of disease. Such impaired host response may be the result of the reduced chemoattraction of lymphocytes, neutrophils and macrophages, and reduced inducible nitric oxide synthase expression (iNOS) and myeloperoxidase (MPO) production in periodontal tissues of p55 TNF-KO mice. Our results demonstrate the mechanisms involved determining periodontal disease severity by TNF-α receptor p55, and its role in providing immune protection to A. actinomycetemcomitans periodontal infection.
Induction of interleukins IL-6 and IL-8 by siRNA
Authors: Pauls E , Senserrich J , Bofill M , Clotet B , Esté J A
Published: November 2006

The HIV-1 co-receptor CCR5 has been thought a relevant target for small interfering RNA (siRNA)-based therapeutics. However, recent findings suggest that siRNA can stimulate innate cytokine responses in mammals. All siRNA agents tested were able to down-regulate the expression of CCR5, albeit with different efficiency (51–74% down-regulation), block HIV-induced syncytia formation between HIV-1 BaL-infected and uninfected CD4+ cells or block single-round HIV-1 infection as measured by a luciferase reporter assay (46–83% inhibition). Conversely, siRNA directed against CCR5 did not affect replication of a vesicular stomatitis virus (VSV) pseudotyped virus, suggesting that inhibition of HIV replication was specific to CCR5 down-regulation. However, two of four siRNA tested were able to induce the production of interleukin (IL) IL-6 (sixfold induction) and IL-8 (ninefold induction) but no interferon (IFN)-α, IFN-β, IFN-γ, tumour necrosis factor (TNF)-α, monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1α, MIP-1β, RANTES, IL-1β, IL-10 or IL-12p70 cytokine induction was noted. In the absence of detectable IFN-α, IL-6 or IL-8 may represent markers of non-specific effects triggered by siRNA.
The involvement of Fc gamma receptor gene polymorphisms in Kawasaki disease
Authors: Biezeveld M , Geissler J , Merkus M , Kuipers I M , Ottenkamp J , Kuijpers T
Published: November 2006

Kawasaki disease is an acute febrile syndrome in infancy, characterized by vasculitis of medium-sized arteries. Without treatment the disease can lead to coronary artery lesions (CAL) in approximately 25% of the children. Therapy consists of intravenous immunoglobulins (IVIG), leading to a decrease of complications to 5–16%. Little is known about the working mechanisms of IVIG. In this study we evaluated the involvement of Fcγ receptors (FcγRs) in Kawasaki disease by the determination of the frequency of known single nucleotide polymorphisms (SNPs) in the genes coding for the FcγRs and compared this with frequencies in a cohort of healthy controls. There was no difference in the distribution of the functionally relevant genotypes for FcγRIIa-131H/R, FcγRIIb-232I/T, FcγRIIIa-158 V/F and FcγRIIIb-NA1/NA2 between the patient group and the healthy controls. Furthermore, there were no polymorphisms linked to the disease severity as indicated by the absence or development of CAL during the disease. Altered transcription or expression of FcγR on specific cell types of the immune system may still play a role in susceptibility and treatment success, but at a level different from the functional SNPs in FcγR genes tested in this study.
Morphometry and digital AgNOR analysis in cytological imprints of benign, borderline and malignant serous ovarian tumours
Authors: Štemberger‐Papić S. , Stanković T. , Vrdoljak‐Mozetič D. , Verša‐Ostojić D. , Krašević M. , Štifter S. , Audy‐Jurković S.
Published: November 2006

Aim:  The aim of the study was to determine values of a quantitative morphometry analysis of nuclear characteristics and argyrophilic nucleolar organizer regions (AgNORs) in differential cytodiagnosis of benign, atypically proliferating (borderline) and malignant serous ovarian tumours.

Methods:  Cytological imprints of benign (n = 20), borderline (n = 19) and malignant (n = 20) ovarian serous tumours were analysed. A computerized, digital analysis was used to determine morphometric nuclear features, the number and characteristics of single AgNORs, cluster AgNORs, total AgNOR and AgNOR area/nucleus (relative area) ratio. According to their size AgNORs were classified in three categories. A one‐way variance analysis and post hoc test (Scheffé) were used for statistical analysis.

Results:  The morphometric nuclear analysis showed that benign, borderline and malignant serous ovarian tumours are statistically different (P < 0.001) according to the area and outline, the values being highest in malignant tumours and lowest in the borderline group. Digital analysis of AgNORs in benign, borderline and malignant groups showed that the total AgNOR number increases with progression of the lesion (meaning tumour malignancy) significantly (P < 0.001) between benign and malignant as well as between borderline and malignant serous ovarian tumours (P < 0.001). The progression of the lesion malignancy was accompanied by a significant (P < 0.001) progressive increase of the total and relative AgNOR area per nucleus. The AgNOR size increases from benign to malignant tumours and a statistically significant difference (P < 0.001) was observed in all three groups regarding small and large AgNORs.

Conclusion:  Combining different markers of morphometric nuclear characteristics and AgNOR values could improve differential cytodiagnosis of benign, borderline and malignant serous ovarian tumours.
Test characteristics of various screening modalities for cervical cancer: a feasibility study to develop an alternative strategy for resource‐limited settings
Authors: Sodhani P. , Gupta S. , Sharma J. K. , Parashari A. , Halder K. , Singh V. , Sehgal A.
Published: November 2006

Objective:  To determine test characteristics – sensitivity, specificity, positive and negative predictive values – of different screening modalities to detect cervical precancerous and cancerous lesions in order to devise an effective alternative strategy for cervical cancer screening in resource‐poor settings.

Methods:  A total of 472 women presenting with nonspecific gynecologic symptoms were screened by cytology, visual inspection with acetic acid application (VIA), VIA with magnification (VIAM) and human papillomavirus (HPV)‐DNA testing. Colposcopic examination was performed in all and on‐site biopsy was taken if any grade I and above lesion was detected on colposcopy (230). On histopathological examination, 105 showed cervical intraepithelial neoplasia II and above lesions. Sensitivity, specificity and predictive values for each test were calculated taking colposcopy and or directed biopsy as the gold standard. Comparisons were made with cytology in order to assess the feasibility of alternative strategies in resource‐poor settings.

Results:  VIA was less sensitive (86.7% versus 91.4%) but more specific (90.7% versus 86.6%) than cytology at low grade squamous intraepithelial lesion (LSIL) threshold but the difference was not statistically significant (P > 0.01). HPV testing improved the sensitivity over cytology (97.1% versus 91.4%) but there was a nonsignificant loss of specificity (84.2% versus 86.6%). Results of VIAM were more or less similar to VIA.

Conclusion:  VIA can be used as a mass screening tool for cervical cancer in resource‐poor settings.
Improving the NHS cervical screening laboratory performance indicators by making allowance for population age, risk and screening interval
Authors: Blanks R. G. , Moss S. M. , Denton K.
Published: November 2006

Objective:  One of the key performance measures in the monitoring of the NHS cervical screening programme is the targeting of laboratories with very high or low percentages (outside the 10th–90th percentile) of adequate smears that have moderate dyskaryosis or worse. These laboratories are assumed to include those laboratories that may have extremes of sensitivity and specificity. A clear limitation with this methodology is that laboratories do not examine smears from women with the same underlying risk, age distribution or screening interval and adjustment for these factors should considerably improve the method.

Methods:  This paper describes a method that allows for these confounding variables and a new age‐risk‐interval adjusted moderate dyskaryosis or worse rate (ARI‐adjusted mod+ rate) can be calculated. The adjusted rate is the rate of moderate or worse dyskaryotic smears that the laboratory would have detected had it been screening women with an English ‘average’ age‐risk‐interval. All laboratories can therefore be compared using this method.

Results:  The methodology is illustrated using data from the NHSCSP South West Region. The particularly low percentage of moderate or worse smears detected by one or two laboratories can be shown to be due to a local screened population with a very low risk because of a high mean age, relatively short screening interval and census variables associated with a low risk, rather than any under‐calling by the associated laboratories.

Conclusions:  The ARI‐adjusted mod+ rate requires to be calculated for all laboratories in England if it is to be used as a primary performance indicator. Alternatively, it can be used to further examine laboratories that are deemed to be outliers using the current methodology.
Interlaboratory reproducibility of atypical glandular cells of undetermined significance: a national survey
Authors: Confortini M. , Di Bonito L. , Carozzi F. , Ghiringhello B. , Montanari G. , Parisio F. , Prandi S. ,   the GISCi Working Group for Cervical Cytology
Published: November 2006

Objective:  The aim of this study was to evaluate the inter‐laboratory reproducibility for atypical glandular cells (AGC) (The Bethesda System (TBS) 2001) of the laboratories involved in the screening programmes in Italy.

Methods:  A set of 35 selected slides were circulated among 167 laboratories involved in local population‐based cervical screening programmes. Each laboratory provided one single diagnosis per smear. The smears were read blind to the original diagnosis and to the diagnoses provided by other laboratories. A ‘majority’ diagnosis was defined for each case and assumed as the reference standard. The diagnosis provided from each laboratory was compared with the majority diagnosis.

Results:  According to the majority report the 35 slides in the set were classified as negative in nine cases, AGC in eight, adenocarcinoma in eight, and squamous lesion or squamous + glandular lesion in 10. The crude agreement between all pairs of laboratories was 49.43%. K‐values were 0.46, 0.21, 0.34, 0.36 and 0.32 for negative, AGC/AIS (adenocarcinoma in situ of endocervix), AdenoCa, Sq/Sq + Gl and all reporting categories respectively. Concordance according to overall K was moderate to substantial in 77% of the participating laboratories.

Conclusions:  The present study shows that the AGC category is not easily reproducible. The data confirmed the importance, in a screening scenario, of AGC/AIS diagnoses, but also presented difficulties in differentiating between the two diagnoses. In addition to the results obtained from the circulation of the slides, laboratories which had annually a low number of cervical smears were able to gain experience focused on particular morphological pictures.
The quality of smear taking training for hospital medical trainees
Authors: Mokate T. , Abidogun K. , Watson A. J. S.
Published: November 2006

Introduction:  Training for cervical smear takers in primary care is well established. We aimed to assess the adequacy of training in cervical smear taking among hospital medical trainees.

Method:  An anonymous questionnaire survey was conducted among hospital medical trainees within the North West region of England. The questionnaire assessed the trainees’ level of confidence in their knowledge and skills in the theoretical and practical aspects of cervical smear taking, and the training they had received. The standards used were as stated in the National Health Service Cervical Screening Programme Resource Pack for Training Smear Takers (July 1998). These included practical smear taking, pre‐ and post‐test counselling, the background to cervical screening, the National Health Service Cervical Screening Programme, the role of the Health Authority and the laboratory, provision for women with special needs, acceptability, accountability and quality assurance.

Results:  Seventy‐eight per cent of trainees returned the questionnaires. Although all the trainees were confident about taking smears, a significant percentage was concerned about the theoretical and specific issues associated with smear taking such as the care of women with special needs. Only 60.7% had received formal training in practical smear taking. Furthermore, only 48.3% had been formally trained in the clinical management of smears.

Conclusion:  There appears to be an urgent need for formal training in the theory and practice of smear taking and for the standardization of the course content. The introduction of training in liquid‐based cytology provides us with a golden opportunity to address this deficit.
Assessing the wellbeing of cytoscreeners: experience in two NHS cytology laboratories
Authors: Dowie R. , Stoykova B. , Desai M.
Published: November 2006

Objective:  Cervical screening programmes in England are in transition as the liquid‐based cytology (LBC) method replaces conventional Papanicolaou screening and staff in NHS laboratories are trained to analyse LBC smears. Cytoscreeners and biomedical scientists undertake routine microscopy of slides, but the scientists usually have a wider professional role. Attitudinal surveys were carried out in laboratories where LBC was partially introduced.

Methods:  Staff in two cytology laboratories in Greater Manchester were surveyed twice over 6 months. The questionnaire assessed work pressures using scales from the Measures of Work Characteristics instrument, work‐related stress using the General Survey version of the Maslach Burnout Inventory, job intentions and job satisfaction.

Results:  Cytoscreeners, many aged over 50 years, formed over 60% of respondents in both surveys (27/42 in the first survey), and biomedical scientists and doctors, 30%. Both groups were under moderate pressure from work demands in each survey, but cytoscreeners had significantly less autonomy over their working methods (P < 0.001). Although both groups experienced similar levels of exhaustion, cytoscreeners were much more cynical or indifferent towards work in the second survey (P = 0.008) and had lower expectations of being effective (P < 0.001). For the cytoscreeners, there were strong negative correlations in both surveys between cynicism and the work characteristics of influencing decisions and autonomy/control.

Conclusions:  The strength of the relationship between work performance and wellbeing serves to emphasize the importance of the new LBC technology in ameliorating low morale where it exists. Further attitudinal research involving larger samples of laboratories is warranted to assess the full impact of this innovation.
Improved endocervical sampling and HPV viral load detection by Cervex‐Brush® Combi
Authors: Depuydt C. E. , Benoy I. H. , Bailleul E. J. , Vandepitte J. , Vereecken A. J. , Bogers J. J.
Published: November 2006

Objective:  Liquid‐based cytology (LBC) for cervical screening is becoming increasingly used. Together with SurePath® LBC, various collecting devices can be utilized, among which the Cervex‐Brush® is the most widely used. The new Rovers® Cervex‐Brush® Combi combines the advantages of the Cervex‐Brush® with the EndoCervex‐Brush® increasing sampling of the endocervical canal. The objective of this study was to analyse and to compare the Cervex‐Brush® Combi with the Cervex‐Brush® for the collection of squamous and endocervical cells, human papillomavirus (HPV) typing/quantification and disease detection in SurePath® LBC.

Methods:  Using either the Cervex‐Brush® or the Cervex‐Brush® Combi 100 consecutive SurePath® LBC samples were collected using each brush type. All 200 slides were read by the FocalPointTM and screened by guided screening using slide wizards. The viral load of HPV type 16 E7, 18 E7, 31 E6, 33 L1, 33 E6, 35 E4, 39 E7, 45 E7, 51 E6, 52 L1, 52 E7, 53 E6, 56 E7, 58 L1, 58 E6, 59 E7, 66 E6 and 68 E7 was determined using a TaqMan‐based real‐time quantitative PCR analysis.

Results:  The mean number of sampled squamous cells did not differ between the two brush types (54 963 versus 54 595 cells). The use of the Cervex‐Brush® Combi, however, resulted in a two‐ to threefold increase in the number of sampled endocervical cells (P < 0.00001). Using the Cervex‐Brush® Combi slightly more lesions were detected (three versus two low‐grade squamous intraepithelial lesions), and resulted in the detection of more atypical squamous cells of undetermined significance (six versus three). In the Cervex‐Brush® group, 60% (3/5) of abnormal smears were positive for oncogenic HPV types, whereas 66.7% (6/9) of abnormal smears in the Cervex‐Brush® Combi group tested positive. The median HPV viral load for samples taken with the Cervex‐Brush® Combi was 0.1825 copies/cell and was significantly higher than in samples taken with the Cervex‐Brush® (0.0042 copies/cell) (P = 0.02).

Conclusion:  Sampling with the Cervex‐Brush® Combi resulted in the collection of the same amount of squamous cells, but in a two to threefold harvest of endocervical cells. This led to the detection of a higher viral load for oncogenic HPV and an increase in the number of detected abnormal smears.
Hosts and symptoms of Plum pox virus: fruiting Prunus species
Authors: Llácer G. , Cambra M.
Published: November 2006

A description of plum pox (sharka) symptoms on European and Japanese plum, apricot, peach, sweet and sour cherry and almond is presented. Plum pox is a serious disease mainly because it produces severe symptoms on fruits of numerous cultivars of Prunus species, causing fruit to fall or making them unfit for consumption.
Causal agent of sharka disease: Plum pox virus genome and function of gene products
Authors: Salvador B. , García J. A. , Simón‐Mateo C.
Published: November 2006

Our understanding of the molecular biology of Plum pox virus (PPV) and of the interactions between the virus and its hosts has advanced notably in recent years. Complete genome sequences have been obtained showing up the existence of new subgroups. Functions related to replication, symptom induction, virus movement, and interference with host defense mechanisms have been assigned to the different protein products derived from the proteolytic processing of PPV polyprotein. Moreover, the use of new approaches to define protein–protein interactions between the different viral proteins and between these and the host proteins has begun to yield the first results improving our understanding of how the virus and its host interact during infection.
Causal agent of sharka disease: historical perspective and current status of Plum pox virus strains
Authors: Candresse T. , Cambra M.
Published: November 2006

Knowledge of the existence of strains or types of Plum pox virus (PPV), defined as monophyletic groups of closely related isolates sharing similar properties, has gradually emerged. Early attempts illustrated both the interest and the limits of biological properties for discrimination of isolates but in the late 1970s gel‐immunodiffusion assays allowed the grouping of PPV isolates into two serogroups typified by the Dideron (D) and Marcus (M) isolates. Technical difficulties, however, limited the widespread use of this approach and only in the early 1990s, with the availability of more discriminating and higher throughput techniques, did it become clear that PPV isolates could be allocated into two major groups. This led to the description of the strains, or types, PPV‐D and PPV‐M. Following this, distant PPV isolates typifying other strains such as El Amar (PPV‐EA), Cherry (PPV‐C) or Winona (PPV‐W) were identified, as well as the widespread group of recombinants known as PPV‐Rec. Each of the six known strains or types can now be identified using a range of serological or molecular techniques. However, their evolutionary history and, in particular, the reason for the wide prevalence of the PPV‐D, ‐M and ‐Rec types relative to PPV‐EA, ‐C or ‐W is still a matter of speculation.
Detection of Plum pox virus: biological methods
Authors: Gentit P.
Published: November 2006

This paper describes biological testing techniques for Plum pox virus (PPV), focussing in particular on woody indicators such as Prunus persica (L.) Batsch GF305 and P. tomentosa (Thunb.) Wall.
Epidemiology of sharka disease in France
Authors: Labonne G. , Dallot S.
Published: November 2006

Plum pox virus was first detected in France in the 1960s. Both PPV‐D and PPV‐M strains are present but epidemics related to the PPV‐M strain detected in the late 1980s are the most problematic. The two PPV strains have unequal distributions in peach and apricot orchards and different prevalences. More than 20 different aphid species have been identified as vectors of PPV but most of them do not colonize Prunus species. Thus, aphids involved in the spread of PPV in orchards are essentially visiting aphids. The main sources of inoculum for the vectors are leaves and fruits of infected stone‐fruit trees. Spontaneous, wild and ornamental Prunus species such as Prunus dulcis, P. spinosa or P. pissardii are susceptible to PPV isolates found in France but their role as a reservoir in sharka epidemics is probably negligible. The disease spreads rapidly in orchards but the rate of progression may vary according to the identity of the PPV strain and the Prunus species. Analysis of spatial patterns of disease has shown that secondary spread by aphids frequently occurs over short distances in the orchards (aggregated patterns) but also that dissemination at longer distances (of several hundred metres) is a common event.
Control and monitoring: phytosanitary situation of Plum pox virus in Australia
Authors: Rodoni B. , Merriman P. , Moran J. , Whattam M.
Published: November 2006

Sharka disease does not occur in Australia and is recognized as the most serious disease threat to Australia's stone fruit industry. Strategies are in place to minimize the risk that exotic diseases like sharka should establish in Australia. These are achieved by implementing: (1) pre‐import measures to reduce the threat of entry; (2) controls at the point of entry and (3) response and contingency planning to contain or eradicate any incursions into Australia. An active and on‐going awareness programme within the Australian stone fruit industry is aimed at early detection of sharka disease in the orchard.
Control and monitoring: Post‐Entry Quarantine System for stone fruits in Chile
Authors: Muñoz M. , Collao M. , Peña X.
Published: November 2006

In Chile a Post‐Entry Quarantine System for propagating plant material has been established to prevent the introduction of quarantine diseases and to maintain good national phytosanitary status. Four types of quarantine facilities are in place and may be used for Prunus spp. and hybrids: absolute quarantine, filter quarantine and farm quarantines levels 1 and 2. All imported Prunus material is submitted to biological, serological and molecular analysis for Plum pox virus (PPV) detection.
Control and monitoring: monitoring and eradication of sharka in south‐east Italy over 15 years
Authors: Myrta A. , Di Terlizzi B. , Savino V. , Martelli G. P.
Published: November 2006

When the first foci of sharka were discovered in Puglia region (south‐east Italy) in the late 1980s, the regional agricultural authorities launched a programme for Plum pox virus (PPV) monitoring and disease eradication. The infecting virus strain was identified as PPV‐D. From 1989 to 1993, a strong eradication campaign was successfully carried out involving 13 plum and 2 apricot orchards with different levels of infection. During 1994–2000, besides plum, apricot and peach, monitoring was extended to sweet cherry. At that time, surveys and testing did not reveal any new PPV focus, but the eradication of infected trees continued in a couple of orchards. In 2001–05, particular attention was paid to peach, as devastating PPV‐M outbreaks had developed in other areas of the country. A new PPV focus was found in apricot, caused by PPV‐Rec, which was promptly eradicated. In the following two years, surveys in the once infected orchard and surrounding peach plantings did not detect any virus spread. The endeavour has taken 15 years making this PPV monitoring and eradication programme the longest in Italy. Its overall results indicate that the fruit tree industry in Puglia region can now be regarded as essentially PPV‐free.
Control and monitoring: eradication of Plum pox virus in Switzerland
Authors: Ramel M. E. , Gugerli P. , Bünter M.
Published: November 2006

In 1967, sharka disease (Plum pox virus; PPV) was detected for the first time in Switzerland. By the end of the 1970s, the disease was considered as eradicated after a rigorous campaign of survey and eradication. Since then, periodical inspections and random testing of imported plants and young orchards have been performed by Swiss plant protection services. This permitted the detection of outbreaks of PPV in 2004. Consequently, a new survey and eradication campaign began.
Control and monitoring: control of Plum pox virus in the United Kingdom
Authors: Mumford R. A.
Published: November 2006

Plum pox virus (PPV) was first identified in the United Kingdom in 1965. Despite a rigorous eradication policy, the disease spread quickly and established itself in all the main plum‐growing areas in England. In 1975, the policy was changed from a blanket eradication campaign to one of containment; retaining statutory control of PPV on propagation material but allowing the industry to control the disease in orchards. As part of the current containment campaign, annual surveys are carried out on propagation material. These surveys show that the incidence of PPV in this material is very low and that only the D‐strain is present. The precise situation regarding PPV incidence in commercial orchards is unknown. Given the low incidence in propagation material, it is likely that PPV is uncommon in actively managed orchards. However, some infected orchards probably do still exist, especially older, unmanaged or abandoned ones. Overall, the history of PPV control in the UK is one of unsuccessful eradication but successful containment. The UK experience demonstrates that given the right combination of strain and host, alongside a regular testing regime, it is possible to control PPV through the establishment of a regulated certification scheme and the supply of virus‐free planting material.
Breeding for resistance: conventional breeding for Plum pox virus resistant apricots (Prunus armeniaca L.) in Greece
Authors: Karayiannis I.
Published: November 2006

A large apricot breeding programme has been conducted at NAGREF‐Pomology Institute, Naoussa‐Greece, for the control of sharka disease, since 1989. Ten apricot cultivars of North American origin: ‘Stark Early Orange’, ‘Stella’, ‘NJA2’, ‘Sunglo’, ‘Veecot’, ‘Harlayne’, ‘Henderson’, ‘Goldrich’, ‘Orangered’ and ‘Early Blush’, selected for their resistance to the highly virulent local strain of Plum pox virus (PPV)‐M (Marcus), have been used as parents in crosses with quality cultivars, mainly with the local cv. Bebecou, from 1989 to 2003. Approximately 7000 hybrids have been created. Resistance to PPV was the main criterion of selection. Most hybrids have been subjected to artificial inoculation by PPV‐M and examined for symptom expression for more than five years. Indexing to GF‐305, as well as laboratory diagnostic tests, have been applied. The genetic analysis showed that: (1) 50% of the hybrids inherited resistance to PPV in the families where cvs. Stark Early Orange, NJA2, Sunglo, Veecot and Harlayne were used as a parent, and (2) 100% of the hybrids inherited resistance to PPV in the families where cv. Stella was one of the parents. Resistance to PPV appears to be under simple genetic control involving one gene locus. Promising apricot selections resistant to PPV‐M have been released.
Breeding for resistance: breeding for resistance to Plum pox virus in Italy
Authors: Bassi D.
Published: November 2006

Plum pox virus (PPV) resistance in apricot and peach in Italy has been researched in several Italian institutions, including the Institutes for Fruit Tree Culture and Plant Pathology in Rome, which are assessing resistance, and the Universities of Bari, Bologna and Milan, examining breeding techniques, both by cross‐breeding and genetic transformation approaches. A reliable protocol for resistance assessment has been set up, in which biological indexing, ELISA and PCR techniques are used. No source of resistance has yet been found in peach. Hybridization with Prunus davidiana could be investigated given the promising results obtained within the INRA programme in France. Almost 6000 apricot seedlings from controlled crosses between resistant parents have been investigated in search of possible molecular markers, but no positive results have been obtained so far, probably due the fact that resistance to both D and M strains is sought. The transformation of cultivars has proven difficult in apricot where the regeneration step is particularly critical, while in peach a reliable regeneration protocol still remains to be set up.
Breeding for resistance: breeding for Plum pox virus resistant apricots (Prunus armeniaca L.) in the Czech Republic
Authors: Krška B. , Salava J. , Polák J.
Published: November 2006

Plum pox virus (PPV) causes serious damage in apricots grown in the Czech Republic and other countries where it is present. To study PPV resistance in apricot, three backcrosses between apricot cultivars or selection resistant to PPV and apricot cultivars or selections susceptible to PPV (‘LE‐3218’ × ‘Stark Early Orange’, ‘LE‐3241’ × ‘Vestar’ and ‘LE‐3246’ × ‘Vestar’) were performed. The seedlings were repeatedly inoculated with PPV‐M strain by an infected chip. The resistance of the plants was evaluated by visual inspection for symptoms and ELISA in three consecutive growth periods. The Chi‐square (χ2) test was used to analyse the data. It was found that two independent dominant complementary genes conditioned PPV resistance in apricot in the case of the backcrosses. The mode of inheritance for resistance to PPV in cv. Harlayne was determined from F1 progenies. Resistance to PPV in ‘Harlayne’ was controlled by three independent complementary dominant genes. This knowledge will help us in effective planning of apricot breeding programmes.
Control of Plum pox virus through the use of genetically modified plants
Authors: Scorza R. , Ravelonandro M.
Published: November 2006

Genetic resistance to Plum pox virus (PPV) is the most viable alternative for long‐term control of sharka disease. In addition to the classical approaches to producing resistant germplasm and cultivars, genetic transformation offers a promising genetic approach to resistance. We show, using the example of C5 plum, that genetically engineered resistance can provide durable, stable and high levels of PPV resistance. A review of the results of work with C5 including molecular analyses of resistance and long‐term field testing is presented.
The use of Plum pox virus as a plant expression vector
Authors: García J. A. , Lucini C. , García B. , Alamillo J. M. , López‐Moya J. J.
Published: November 2006

Among the biotechnological uses of plant viruses, the expression of foreign sequences through virus‐based vectors represents a promising research area. The potyvirus Plum pox virus (PPV) has been used to design expression vectors which have allowed successful expression of foreign sequences in plants, either in the form of small peptides fused to the viral coat protein, or as whole independent proteins inserted on different points of the genome. The present review describes the different PPV‐based vectors that have been produced, including information regarding relevant aspects of their use, such as the optimal location of peptides or the stability of inserts. Recent developments, like the expression of proteins on stone fruit trees by using PPV‐based vectors capable to infect woody plants, are also described.
Phytophthora kernoviae and P. ramorum: host susceptibility and sporulation potential on foliage of susceptible trees1
Authors: Denman S. , Kirk S. , Whybrow A. , Orton E. , Webber J. F.
Published: November 2006

Phytophthora kernoviae and P. ramorum are introduced, invasive pathogens in the UK. Both species are adapted for aerial dispersion and have a wide host range, many of which are common to both pathogens. The diseases they cause are foliar necrosis and shoot tip dieback on both tree and ornamental hosts, and bleeding cankers on tree hosts. Inoculum is produced on infected foliage but not on bleeding cankers in both cases. Proactive measures to prevent disease spread and to evaluate the risks posed by these pathogens are being undertaken. Amongst others, these include using the detached leaf assay to get an indication of tree foliage susceptibility, and inoculating wounded stems of saplings to get an idea of under‐canopy sapling and nursery tree susceptibility. The sporulation potential on selected susceptible hosts was assessed, and finally, surveys which are still ongoing were carried out in south‐west England. Results of this work are presented and discussed.
Diagnosis of Phytophthora ramorum– evaluation of testing methods1
Authors: Lane C. R. , Beales P. A. , Hughes K. J. D. , Tomlinson J. , Boonham N.
Published: November 2006

Phytophthora ramorum is a recently described pathogen that has resulted in the introduction of emergency European Community phytosanitary legislation in November 2002. The Department of Environment, Food and Rural Affairs Plant Health and Seeds Inspectorate (PHSI) initiated a survey in England and Wales in autumn 2001 to ascertain whether the organism was present. In April 2002, P. ramorum was found for the first time in the UK and for the first time in Europe since the initial reports in the Netherlands and Germany. Following this finding, P. ramorum has been reported mainly on rhododendrons and viburnums but also on a wide range of other ornamental plants in both the UK and other European countries. Effective disease management and implementation of plant health legislation is reliant upon rapid and accurate disease diagnosis based upon recognition of symptoms in the field and identification of the cause by testing. Other diseases and disorders prevent reliable identification of the problem at the time of inspection with typically only 20–30% of samples submitted testing positive for Phytophthora species. This paper describes the evaluation of a direct real‐time PCR and a rapid serological assay (lateral flow device) developed by CSL for testing for P. ramorum and Phytophthora spp., respectively.
Surveys for Phytophthora ramorum in the Czech Republic1
Authors: Běhalová M.
Published: November 2006

The State Phytosanitary Administration (SPA) of the Czech Republic has been conducting an official survey for Phytophthora ramorum since 2003. A single outbreak of P. ramorum was detected by the SPA on guelder rose (Viburnum bodnantense) in autumn 2003 during a phytosanitary inspection in an ornamental nursery made on the request of owner of the nursery as he had noticed plants of Viburnum bodnantense withering. The infested lot consisted of 15 plants, which were imported from Italy into the Czech Republic at the end of April in 2003. A post‐entry inspection was carried out at the place of destination just after arrival and no visual symptoms were seen at that time. The lot was destroyed by the owner of the nursery immediately after the sampling. A survey was carried out by the SPA in all nurseries in which host plants coming from the same origin as the infested plants had been planted. No other outbreak was found. Phytosanitary inspections were carried out in ornamental nurseries and garden centres, public parks and forestry sites. No outbreak was detected in 2004 (451 visual inspections) and 2005 (522 visual inspections). The status of Phytophthora ramorum in the Czech Republic is: absent, one isolated outbreak linked to an imported consignment was eradicated.
Project management of an eradication campaign1
Authors: Ward M. G.
Published: November 2006

Applying project management to a task helps to define objectives, resources, timescales, risks and governance arrangements. It can secure commitment of effort and expertise from other organisations, and from other parts of a large organisation, and ensure effective co‐ordination. The UK plant health services are increasingly using project management disciplines and terminology. To direct eradication and containment action against Phytophthora ramorum and Phytophthora kernoviae, a programme board has been established on which three government departments and several different fields of expertise are represented. The board has in turn set up subgroups, accountable to it for specific areas of activity. These comprise a science subgroup; a press and publicity subgroup; a field implementation subgroup; and two geographically based subgroups for the areas with Phytophthora kernoviae outbreaks – Cornwall and South Wales. There is also an industry liaison group comprising representatives from trade associations and providing a regular channel of communication with the nurserymen, retailers and woodland and garden owners most directly affected by measures against these pathogens.
Validation of a real‐time PCR method for the detection of Phytophthora ramorum1
Authors: Chandelier A. , Ivors K. , Garbelotto M. , Zini J. , Laurent F. , Cavelier M.
Published: November 2006

To validate a real‐time PCR method for the detection of Phytophthora ramorum, an intra‐laboratory procedure was developed. The specificity of the TaqMan probe/primer sets was determined by carrying out real‐time PCR on total DNA extracted from pure culture of several Phytophthora species. The limit of detection and the potential effects of plant substrates were evaluated by conducting the test on total DNA from healthy plant materials (Rhododendron spp., Viburnum spp. and Pieris spp.) spiked with known amounts of P. ramorum genomic DNA. The PCR efficiency was estimated through the linear regression of the dilution curve. Precision of the TaqMan assay was assessed on material from a single artificially infected plant (Rhododendron spp.). Two kinds of tissues were tested: a severely infected twig and an apparently healthy leaf. Intra‐assay repeatability was evaluated on 10 replicates of the same DNA sample analysed in a single assay. Inter‐assay reproducibility was evaluated on the same DNA sample amplified over five separate assays while the intersample reproducibility was evaluated on separate DNA extractions of four samples from both plant tissues amplified in a single assay.