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INTRODUCTION: Up to 39% of women in Denmark do not participate regularly in the cervical cancer screening program and initiatives to increase participation are called upon. The primary aim of this study was to describe previous screening history and characteristics of women attending screening in a walk-in clinic. Furthermore, we wanted to investigate barriers to cervical cancer screening.MATERIAL AND METHODS: We designed a walk-in clinic that was open 2 days a week from 16.00 to 19.00 h, located in the Departments of Gynecology in the two main hospitals of the North Denmark Region. The main purpose of the clinic was cervical cancer screening and the study period was 5 months. Women who were not eligible for screening or had other health complaints were referred to their general practitioner. The women included in the study, filled out a questionnaire regarding educational and occupational status; their screening history was registered using data from the Danish Pathology Register.RESULTS: During the study period, 255 women visited the walk-in clinic. The final study population consisted of 249 women who met the inclusion criteria. Age range of participants was 23-77 years, with a median age of 45 years. The majority of the participants were currently employed (81%) or students (10%), the remaining being retired (5%) or unemployed (4%). Screening history showed that 138 (55.4%) of the women were on time for the screening or delayed less than 6 months compared to their recommended screening interval. Sixty-one women (24.5%) were delayed >6 months but CONCLUSIONS: Women attending the walk-in clinic tended to be primarily actively working or students (91%). All age groups in the screening population were represented. Screening history showed that 44.6% had not followed the recommended screening program.

AIM: In countries like Denmark, cervical cancer incidence is at present relatively high in elderly women, while routine screening stops at age 65 years. On this background, all women aged 69 and above were invited to human papillomavirus (HPV)-screening in Denmark in 2017.METHODS: Women were identified from the Central Population Register and personally invited by digital or ordinary mail to have a screening sample taken by their general practitioner. In four regions, samples were tested for high risk (HPV) with the cobas 4800® HPV-assay, and in the last region with the BD Onclarity® HPV-assay. Participation rate, prevalence of high risk HPV, and proportion of positive samples with HPV16, HPV18, and other high risk HPV-types were tabulated by 5-year age-groups.RESULTS: 455,612 women were invited, and 30.2% (95 confidence interval (CI) 30.0-30.3) participated. Average age of participants was 74.6 years. Overall, 4.3% (95% CI 4.1-4.4) of participants were HPV-positive, of whom 24% had HPV 16/18. HPV-prevalence decreased slightly from 4.5% in women aged 69-73 years to 3.1% in women aged 84-88 years, but was 5.2% in the very small group of participants aged 89+ years.CONCLUSION: Invitation to HPV-screening was well received by elderly women. The HPV-prevalence decreased slightly with increasing age. No rebound of HPV-prevalence after menopause was found when our data were combined with previously published Danish data from younger women. The presently relatively high cervical cancer incidence in elderly women was not reflected in the HPV-prevalence.

To achieve an appropriate and efficient sample in a surveillance program, the goals of the program should drive a careful consideration of the selection method or combination of selection methods to be applied. Therefore, the ongoing analysis and assessment of a surveillance system may include an assessment of the ability of the applied selection methods to generate an appropriate sample. There may be opinions from many technical experts (TEs) and many criteria to consider in a surveillance system so there is a need for methods to combine knowledge, priorities and preferences from a group of TEs. This paper proposes a modified weighted and unweighted TOPSIS (Technique for Order of Preference by Similarity to Ideal Solution) analysis to choose selection methods in surveillance. An example from the Canadian Notifiable Avian Influenza surveillance (CanNAISS) is used to illustrate the method as this surveillance offers unique data with multiple selection methods and subpopulations. The primary objective was to assess the performance of the different selection methods applied in CanNAISS, from 2008 to 2013, in three subpopulations (A-C). A modified TOPSIS (weighted and unweighted) analyses is proposed to aggregate preferences from three TEs and to identify the selection method that was closest to the ideal solution agreed upon by the TEs. Criteria weights were provided individually by three TEs. For the group decision making, internal and external aggregation approaches were used with arithmetic and geometric means. The results of the weighted modified TOPSIS analysis showed that the selection methods that used farm registries yielded high estimates of the relative closeness to ideal-solution. The ranking of selection methods based on the modified TOPSIS weighted analysis, conducted at the individual and group decision making levels were similar. Regardless of the aggregation approach used (internal or external) in group decision making, the use of the arithmetic and geometric means yielded similar estimates of relative closeness to ideal-solution. The unweighted modified TOPSIS analysis yielded similar estimates of the relative closeness to the ideal-solution and therefore making the interpretation of the results difficult. The weighted modified TOPSIS analysis contributed to an informed decision on the best selection method to apply in CanNAISS. The weighted modified TOPSIS analysis is a straightforward and suitable technique to address decision making problems where the profile of the ideal and non-ideal solutions is known a priori by the decision makers.

OBJECTIVE: Cytology findings of atypical squamous cells of unknown significance (ASCUS) or low-grade squamous intraepithelial lesion (LSIL) are common among women under 30, but evidence on best management strategy is insufficient. We therefore investigated how different management strategies used in Denmark influenced biopsy rates and detection of cervical intraepithelial neoplasia (CIN).METHODS: Register-based cohort study including Danish women aged less than 30 years and born 1980-95, with ASCUS/LSIL as their first abnormal cervical cytology in 2008-16. Rates and relative risks (RR) of biopsy and detection of CIN3+, CIN2 and RESULTS: 19,946 women with ASCUS and 19,825 with LSIL were included in the study of whom 92% had adequate information about follow-up. Among women referred directly to biopsy, CIN3+ was detected among 21%, CIN2 in 17%, while 62% had CONCLUSIONS: The choice of management strategy influences both the detection rate for severe lesions (CIN3+) and the proportion of women followed up for potentially insignificant findings.

Vaccination against human papillomavirus (HPV) has been introduced as a public health initiative in many countries, including Denmark since October 2008. It is important to monitor postimplementation effectiveness of HPV‐vaccination at the population‐level. We studied HPV‐prevalence after first invitation to screening at age 23 years in women offered the quadrivalent HPV‐vaccine at the age of 14 years. Randomly selected screening samples from women born in 1994 in four out of five Danish regions were subjected to analysis for HPV in addition to routine cytology. Cobas4800 was used in all participating pathology departments. Data from a Danish prevaccination cross‐sectional study using Hybrid Capture 2, and a Danish split‐sample study using Cobas4800 were used for comparison. In the period from February 2017 to April 2019, 6233 screening samples from women born in 1994 were selected for HPV‐analysis; 27 samples had no HPV‐test and 3 samples had no HPV‐diagnosis, leaving 6203 samples with an HPV‐diagnosis. Prevalence of any high‐risk (HR) HPV was 35%; only 0.9% were positive for vaccine HPV types 16/18 while the remaining 34% were positive for other HR HPV. When comparing with prevaccination prevalence data, HPV‐16/18 decreased by 95%; RR = 0.05 (95% CI 0.04‐0.06), while other HR HPV remained fairly constant; RR = 0.88 (95% CI 0.82‐0.94) and RR = 0.95 (95% CI 0.88‐1.03), respectively. One‐third of women vaccinated as girls with the quadrivalent HPV‐vaccine were HR HPV‐positive at time of first invitation to screening. Vaccine HPV‐types 16 and 18 were almost eliminated, while the prevalence of nonvaccine HR HPV‐types remained constant., What's new? Vaccination against human papillomavirus (HPV) has been introduced as a public health initiative in many countries. However, data on HPV infection prevalence at screening age in HPV‐vaccinated birth cohorts remain scarce. This study found that HPV‐16 and 18 were almost eliminated in a population aged 23‐24 years where 92% of women received quadrivalent HPV vaccination at age 14. Prevalence of HPV 16 and 18 was lower in non‐vaccinated women than in previous birth cohorts not offered HPV‐vaccination. Nonetheless, one third of vaccinated women were positive for high‐risk HPV other than 16/18, with potential implications for cervical cancer prevention and screening programs.

IntroductionThe first birth cohorts of women offered human papillomavirus (HPV) vaccination as girls are now entering cervical screening. However, there is no international consensus on how to screen HPV vaccinated women. These women are better protected against cervical cancer and could therefore be offered less intensive screening. Primary HPV testing is more sensitive than cytology, allowing for a longer screening interval. The aim of Trial23 is to investigate if primary HPV testing with cytology triage of HPV positive samples is a reasonable screening scheme for women offered HPV vaccination as girls.MethodsTrial23 is a method study embedded in the existing cervical screening programme in four out of five Danish regions. Without affecting the screening programme, women born in 1994 are randomised to present screening with liquid-based cytology every third year (present programme arm) or present screening plus an HPV test (HPV arm). The study started 1 February 2017 and will run over three screening rounds corresponding to 7–8 years.AnalysesThe primary endpoint is cervical intraepithelial neoplasia grade 3 or above. The trial is undertaken as a non-inferiority study including intention-to-treat and per-protocol analyses. The potential effect of primary HPV screening with a 6-year interval will be calculated from the observed data.Ethics and disseminationThe study protocol has been submitted to the ethical committee and deemed a method study. All women are screened according to routine guidelines. The study will contribute new evidence on the future screening of HPV vaccinated birth cohorts of women. All results will be published in open-access journal.Trial registrationNCT03049553.

"Freedom from animal disease" is an ambiguous concept that may have a different meaning in trade and science. For trade alone, there are different levels of freedom from OIE listed diseases. A country can: be recognized by OIE to be "officially free"; self-declare freedom, with no official recognition by the OIE; or report animal disease as absent (no occurrence) in six-monthly reports. In science, we apply scenario tree models to calculate the probability of a population being free from disease at a given prevalence to provide evidence of freedom from animal disease. Here, we link science with application by describing how a scenario tree model may contribute to a country's claim of freedom from animal disease. We combine the idea of a standardized presentation of scenario tree models for disease freedom and having a similar model for two different animal diseases to suggest that a simple generic model may help veterinary authorities to build and evaluate scenario tree models for disease freedom. Here, we aim to develop a generic scenario tree model for disease freedom that is: animal species specific, population specific, and has a simple structure. The specific objectives were: to explore the levels of freedom described in the OIE Terrestrial Animal Health Code; to describe how scenario tree models may contribute to a country's claim of freedom from animal disease; and to present a generic swine scenario tree model for disease freedom in Canada's domestic (commercial) swine applied to Aujeszky's disease (AD). In particular, to explore how historical survey data, and data mining may affect the probability of freedom and to explore different sampling strategies. Finally, to frame the generic scenario tree model in the context of Canada's claim of freedom from AD. We found that scenario tree models are useful to support a country's claim of freedom either as "recognized officially free" or as part of a self-declaration but the models should not stand alone in a claim. The generic AD scenario tree model demonstrated the benefit of combining three sources of surveillance data and helped to design the surveillance for the next year. The generic AD scenario model is one piece in Canada's self-declaration of freedom from AD. The model is strongly supported by the fact that AD has never been detected in Canada.

INTRODUCTION: Despite an intensive screening activity, the incidence of cervical cancer in Denmark has remained stable for the last 15 years, while regional differences have increased. To search for explanations, we investigated possible weaknesses in the screening program.MATERIAL AND METHODS: Data on the screen-targeted women were retrieved from Statistics Denmark. Data on screening activity were retrieved from the annual reports from 2009 to 2015 on quality of cervical screening. Coverage was calculated as proportion of screen-targeted women with at least one cytology sample within recommended time intervals. Insufficient follow-up was calculated as proportion of abnormal and unsatisfactory samples not followed up within recommended time intervals. Diagnostic distribution was calculated for samples with a satisfactory cytology diagnosis.RESULTS: Coverage remained stable at 75%-76% during the study period. Annually, approximately 100,000 women are screened before they are eligible for invitation, and 600,000 invitations and reminders are issued resulting in screening of 200,000 women. In 2009, 21% of abnormal and unsatisfactory samples were not followed up within the recommended time interval; a proportion that had decreased to 15% in 2015. Overall, 11% of satisfactory samples with a cytology diagnosis were abnormal, but with surprising variation from 6% to 15% across regions.DISCUSSION: The success of a screening program depends first of all on coverage and timely follow-up of abnormal findings. Our analysis indicated that the currently high incidence of cervical cancer in Denmark may partly be due to low screening coverage. Also worrisome is a high proportion of non-timely follow-up of abnormal findings. Innovative ways to improve coverage and follow-up are urgently needed.

This manuscript was written in honour of Dr. Preben Willeberg to illustrate some tools that may be useful to "make surveillance happen" and to build bridges between science and application in animal health surveillance. The paper illustrates how four elements (science, project management, communication and documentation) may be combined with some tools into a practical framework for Foreign Animal Disease surveillance. Specifically, I will show how the four elements are essential to make surveillance happen and provide a link between science and application in animal health surveillance. The Canadian Notifiable Avian Influenza Surveillance system is used as an example. In surveillance for Foreign Animal Diseases (FAD), with veterinary authorities as the lead and with many stakeholders, project management can support science in building evidence (documentation) that can be delivered (communicated) to trade negotiators and international trade partners. To bridge the gap between science and application in FAD surveillance I propose that we need people with skills in science, project management, communication and documentation.

In 2008, Canada designed and implemented the Canadian Notifiable Avian Influenza Surveillance System (CanNAISS) with six surveillance activities in a phased-in approach. CanNAISS was a surveillance system because it had more than one surveillance activity or component in 2008: passive surveillance; pre-slaughter surveillance; and voluntary enhanced notifiable avian influenza surveillance. Our objectives were to give a short overview of two active surveillance components in CanNAISS; describe the CanNAISS scenario tree model and its application to estimation of probability of populations being free of NAI virus infection and sample size determination. Our data from the pre-slaughter surveillance component included diagnostic test results from 6296 serum samples representing 601 commercial chicken and turkey farms collected from 25 August 2008 to 29 January 2009. In addition, we included data from a sub-population of farms with high biosecurity standards: 36,164 samples from 55 farms sampled repeatedly over the 24 months study period from January 2007 to December 2008. All submissions were negative for Notifiable Avian Influenza (NAI) virus infection. We developed the CanNAISS scenario tree model, so that it will estimate the surveillance component sensitivity and the probability of a population being free of NAI at the 0.01 farm-level and 0.3 within-farm-level prevalences. We propose that a general model, such as the CanNAISS scenario tree model, may have a broader application than more detailed models that require disease specific input parameters, such as relative risk estimates.

The CHD4 helicase is identified as a new component of the genome surveillance machinery in a proteomic screen for factors enriched on chromatin after ionizing radiation (see also related paper by Smeenk et al. in this issue)., In response to ionizing radiation (IR), cells delay cell cycle progression and activate DNA repair. Both processes are vital for genome integrity, but the mechanisms involved in their coordination are not fully understood. In a mass spectrometry screen, we identified the adenosine triphosphate–dependent chromatin-remodeling protein CHD4 (chromodomain helicase DNA-binding protein 4) as a factor that becomes transiently immobilized on chromatin after IR. Knockdown of CHD4 triggers enhanced Cdc25A degradation and p21Cip1 accumulation, which lead to more pronounced cyclin-dependent kinase inhibition and extended cell cycle delay. At DNA double-strand breaks, depletion of CHD4 disrupts the chromatin response at the level of the RNF168 ubiquitin ligase, which in turn impairs local ubiquitylation and BRCA1 assembly. These cell cycle and chromatin defects are accompanied by elevated spontaneous and IR-induced DNA breakage, reduced efficiency of DNA repair, and decreased clonogenic survival. Thus, CHD4 emerges as a novel genome caretaker and a factor that facilitates both checkpoint signaling and repair events after DNA damage.

Purpose To examine the applicability of US reference data from the Pediatric Evaluation of Disability Inventory (PEDI) in a random Danish sample and to assess intrarespondent reliability. Methods The PEDI was administered as a mailed questionnaire that was completed by the parents. Linear regression models and comparisons of item difficulty were performed in the data analysis. Intraclass correlation coefficients were used in the intrarespondent analysis. Results US and Danish children differed on 3 of the 6 subscales of the PEDI. Danish children had significantly lower scores on the caregiver assistance subscales (self-care and mobility) and significantly higher scores on the functional skills subscale (social function). Item difficulty was found to be comparable (r = 0.82-0.97) on all 6 subscales. Intrarespondent reliability was good (intraclass correlation coefficient = 0.62-0.97). Conclusion The PEDI reference values seem partly applicable in a Danish population. In a Danish rehabilitation setting, interpreting PEDI scores with regard to detecting functional delays must be done cautiously.

Three novel triterpenes, (11R,20R)-11,20-dihydroxy-24-dammaren-3-one (1), (17S,20R,24R)-17,25-dihydroxy-20,24-epoxy-14(18)-malabaricen-3-one (2), and (17R,20S,24R)-17,25-dihydroxy-20,24-epoxy-14(18)-malabaricen-3-one (3), were isolated from leaves of Caloncoba echinata. The structures were established using mainly 800 MHz NOESY and HMBC connectivities. The absolute stereochemistry of C-11 in 1 and that of C-17 in 2 were established by the Mosher method. The stereochemistry of the side chains of the malabaricanes is compatible with their biosynthesis by a cascade opening of diepoxides. The isolated triterpenes inhibited growth of Plasmodium falciparum parasites in vitro apparently via incorporation into erythrocyte membrane, as suggested by transformation of erythrocytes into stomatocytes at a concentration level at which the growth inhibition was observed.

Two known phenanthroindolizidine alkaloids, (-)-(R)-13aalpha-antofine (1) and (-)-(R)-13aalpha-6-O-desmethylantofine (2), and two new natural products, (-)-(R)-13aalpha-secoantofine (3) and (-)-(R)-13aalpha-6-O-desmethylsecoantofine (4), were isolated from Cynanchum vincetoxicum. The structures of all compounds were established by means of NMR methods including COSY, NOESY, HSQC, and HMBC experiments, supported by HRMS and optical rotation data. Cytotoxic activity of the isolated alkaloids, and of three other alkaloids previously isolated from Tylophora tanakae, (-)-(R)-13aalpha-tylophorine (5), (-)-(R)-13aalpha-7-O-desmethyltylophorine (6), and (+)-(S)-13abeta-isotylocrebrine (7), was assessed in vitro using a drug-sensitive KB-3-1 and a multidrug-resistant KB-V1 cancer cell line. Structure-activity relationships in this series of alkaloids are discussed. The IC(50) values of some of the alkaloids are in the low nanomolar range, being thus comparable to the activity of clinically used cytotoxic drugs. Previously reported adverse side effects of these alkaloids could possibly be overcome by modern tissue-specific drug targeting techniques.

Nineteen species of Passiflora (Passifloraceae) were examined for the presence of cyanogenic glycosides. Passibiflorin, a bisglycoside containing the 6-deoxy-β- d -gulopyranosyl residue, was isolated from P. apetala , P. biflora , P. cuneata , P. indecora , P. murucuja and P. perfoliata . In some cases this glycoside co-occurs with simple β- d -glucopyranosides: tetraphyllin A, deidaclin, tetraphyllin B, volkenin, epivolkenin and taraktophyllin. P. citrina contains passicapsin, a rare glycoside with the 2,6-dideoxy-β- d - xylo -hexopyranosyl moiety, while P. herbertiana contains tetraphyllin A, deidaclin, epivolkenin and taraktophyllin, P. discophora tetraphyllin B and volkenin, and P. × violacea tetraphyllin B sulfate. The remaining species were noncyanogenic. The glycosides were identified by 1 H and 13 C NMR spectroscopy following isolation by reversed-phase preparative HPLC. From P. guatemalensis , a new glucoside named passiguatemalin was isolated and identified as a 1-(β- d -glucopyranosyloxy)-2,3-dihydroxycyclopentane-1-carbonitrile. An isomeric glycoside was prepared by catalytic hydrogenation of gynocardin. α-Hydroxyamides corresponding to the cyanogenic glycosides were isolated from several Passiflora species. These α-hydroxyamides, presumably formed during processing of the plant material, behave as cyanogenic compounds when treated with commercial Helix pomatia crude enzyme preparation. Thus, the enzyme preparation appears to contain an amide dehydratase, which converts α-hydroxyamides to cyanohydrins that liberate cyanide; this finding is of interest in connection with analysis of plant tissues and extracts using Helix pomatia enzymes.

The study objectives were (1) to conduct a systematic review of the performance of capture–recapture methods; (2) to use empirical data to estimate population size in a small-sized population (turkey breeder farms) and a medium-sized population (meat turkey farms) by applying two-source capture–recapture methods (the Lincoln–Petersen, the Chapman, and Chao's lower-bound estimators) and multi-source capture–recapture methods (the log-linear modeling and sample coverage approaches); and (3) to compare the performance of these methods in predicting the true population sizes (2007 data). Our set-up was unique in that we knew the population sizes for turkey breeder farms (99) and meat turkey farms (592) in Canada in 2007, which we applied as our true population sizes, and had surveillance data from the Canadian Notifiable Avian Influenza Surveillance System (2008–2012). We defined each calendar year of sampling as a data source. We confirmed that the two-source capture–recapture methods were sensitive to the violation of the local independence assumption. The log-linear modeling and sample coverage approaches yielded estimates that were closer to the true population sizes than were the estimates provided by the two-source methods for both populations. The performance of both multi-source capture–recapture methods depended on the number of data sources analyzed and the size of the population. Simulation studies are recommended to better understand the limits of each multi-source capture–recapture method.

The objective of the present study was to analyze the role of the mesothelial markers desmin and N-cadherin in the diagnostic panel of serous effusions. A total of 181 pleural and peritoneal effusions consisted of 101 cases cytologically diagnosed as malignant (89 carcinomas, 12 mesotheliomas), 78 benign, and 2 inconclusive specimens. All specimens were immunostained using 11 antibodies, against epithelial membrane antigen, Ber-EP4, carcinoembryonic antigen, E-cadherin, CA 125, N-cadherin, desmin, calretinin, p53, vimentin, and CD45. After evaluation of immunocytochemistry results, 110 specimens were diagnosed as malignant (98 carcinomas, 12 mesotheliomas) and 71 as benign (56 cellular, 15 paucicellular). The presence of desmin was detected in benign mesothelial cells in 47 of 56 (84%) reactive cellular specimens compared with 1 of 12 (8%) malignant mesotheliomas and 2 of 98 (2%) carcinomas. N-cadherin was expressed in 48 of 56 (86%) reactive cases, 12 of 12 (100%) mesotheliomas, and 47 of 98 (48%) carcinomas. In carcinomas, N-cadherin expression was most often seen in ovarian carcinoma but was also found in other carcinomas. Calretinin, an established marker of mesothelial cells, was detected in 52 of 56 (93%) reactive specimens, 11 of 12 (93%) mesotheliomas, and 3 of 98 (3%) carcinomas. Evaluation of staining results led to reclassification of six malignant specimens as benign, whereas 17 cases diagnosed as benign and the two diagnosed as inconclusive were classified as malignant. In conclusion, desmin appears to be a promising marker for the distinction between reactive mesothelium and malignant epithelial cells in terms of both specificity and sensitivity, and its complementary use with calretinin is recommended. Unlike calretinin, it may also prove valuable for the distinction between benign and malignant mesothelial cells. N-cadherin does not have a role in the distinction between mesothelial and epithelial cells. However, it may prove useful in the characterization of carcinomas of unknown origin. As has previously been shown, a significant number of diagnoses that are based on morphologic examination alone are modified after the use of a broad antibody panel.

Scenario tree models with temporal discounting have been applied in four continents to support claims of freedom from animal disease. Recently, a second (new) model was developed for the same population and disease. This is a natural development because surveillance is a dynamic process that needs to adapt to changing circumstances - the difficulty is the justification for, documentation of, presentation of and the acceptance of the changes. Our objective was to propose a systematic approach to present changes to an existing scenario tree model for freedom from disease. We used the example of how we adapted the deterministic Canadian Notifiable Avian Influenza scenario tree model published in 2011 to a stochastic scenario tree model where the definition of sub-populations and the estimation of probability of introduction of the pathogen were modified. We found that the standardized approach by Vanderstichel et al. (2013) with modifications provided a systematic approach to make and present changes to an existing scenario tree model. We believe that the new 2013 CanNAISS scenario tree model is a better model than the 2011 model because the 2013 model included more surveillance data. In particular, the new data on Notifiable Avian Influenza in Canada from the last 5 years were used to improve input parameters and model structure.

This paper discusses statistical modelling for data with a hierarchical structure, and distinguishes in this context between three different meanings of the term hierarchical model: to account for clustering, to investigate variability and separate predictive equations at different hierarchical levels (multi-level analysis), and in a Bayesian framework to involve multiple layers of data or prior information. Within each of these areas, the paper reviews both past developments and the present state, and offers indications of future directions. In a worked example, previously reported data on piglet lameness are reanalyzed with multi-level methodology for survival analysis, leading to new insights into the data structure and predictor effects. In our view, hierarchical models of all three types discussed have much to offer for data analysis in veterinary epidemiology and other disciplines.

Freedom from infection modeling, using scenario trees, has become an established methodology and is well described in the literature. However, standards for organizing and reporting the surveillance information incorporated into such models are less developed. Canada has been routinely testing for Trichinella spiralis in market hogs in federally inspected slaughter plants since the late 1990s. By way of presenting our work on T. spiralis in Canadian hogs, we propose that information in surveillance models be organized in distinct categories, each with specific parameters and values that are thoroughly described and justified. The proposed categories are: (1) definitions for the objectives, (2) initial time period, (3) inputs, (4) data, (5) model settings, (6) outputs, and (7) validation. Having a standardized manner of reporting such studies will facilitate their validation and expedite their evaluation by experts in the field and their use in trade negotiations.

Dehydroabietinol isolated from Hyptis suaveolens (L.) Poit. was found to inhibit growth of chloroquine-sensitive as well as chloroquine-resistant strains of Plasmodium falciparum cultivated in erythrocytes in vitro (IC 50 26-27 microM). However, erythrocytes exposed to dehydroabietinol were transformed in a dose-dependent manner towards spherostomatocytic forms with concomitant formation of endovesicles, as disclosed by transmission electron microscopy. The erythrocyte shape alterations caused by dehydroabietinol correlated well with its apparent IC 50 value. Thus, dehydroabietinol incorporates into the erythrocyte membrane, and since invasion and survival of Plasmodium parasites is known to depend on the function of the erythrocyte membrane, the observed antiplasmodial effect of dehydroabietinol is presumably an indirect effect on the host cell. Because of these findings, microscopic investigations should be generally used to support claims of antimalarial effects of apolar natural products.

[structure: see text] Leaves of the edible passion fruit plant, Passiflora edulis, contain benzylic beta-D-allopyranosides 1 and 2, representatives of a rare class of natural glycosides with D-allose as the only sugar constituent. The glycoside 1 is the first known cyanogenic glycoside containing a sugar different from D-glucose attached directly to the cyanohydrin center. Asymmetric perturbation of the (1)L(b) transition of the benzene chromophore was shown to be useful for determination of absolute configuration of the cyanohydrin center of aromatic cyanogenic glycosides.

The Pediatric Evaluation of Disability Inventory (PEDI) assesses functional status in children with disabilities aged 0.5-7.5 years. The purpose of this study was to examine if the Danish version of the PEDI was able to discriminate between nondisabled children and children with cerebral palsy (CP) or juvenile idiopathic arthritis (JIA). Convenience sampling was used to select the clinical samples that consisted of 22 children with CP and 14 children with JIA aged 1.5-7.5 years. An additional sample, consisting of 224 nondisabled children aged 1.0-5.9 years, was obtained on the basis of random sampling. The PEDI was administered as a questionnaire completed by the parents. Results from receiver operating characteristics (ROC) analysis showed that in the case of discrimination between children with CP and nondisabled children, the sensitivity and specificity were 77.3 and 97.8%, respectively. In the discrimination between children with JIA and nondisabled children, the sensitivity and specificity were 99.8 and 81.7%, respectively. Study results indicate that the Danish version of the PEDI shows good discriminative validity. Further studies of the discriminative validity are recommended.

Two previously known phenanthroindolizidine alkaloids, (-)-10beta-antofine N-oxide (1) and (-)-10beta, 13aalpha-14beta-hydroxyantofine N-oxide (2), and a novel alkaloid, (-)-10beta,13aalpha-secoantofine N-oxide (3), were isolated from aerial parts of Cynanchum vincetoxicum. Their structures were established by means of NMR methods, including COSY, NOESY, HSQC, and HMBC experiments, as well as from their CD spectra. Cytotoxic activity of the alkaloids was assessed in vitro using both a drug-sensitive KB-3-1 and a multi-drug-resistant KB-V1 cancer cell line. The antofine derivatives (1 and 2) showed pronounced cytotoxicity against the drug-sensitive cell line (IC(50) values about 100 nM), whereas the secoantofine derivative (3) was considerably less active. The KB-V1 cell line showed a marginal resistance against all alkaloids, demonstrating that these compounds are poor substrates for the P-glycoprotein (P-170) efflux pump.

Swine influenza (SI) is a disease of significance for the swine industry, and vaccination is often recommended as a way to reduce its impact on production. The efficacy of SI vaccines is well established under experimental conditions, but information about field efficacy is scarce. The objective of this study was to evaluate the efficacy of a commercial inactivated bivalent (H1N1/H3N2) vaccine under conditions of natural exposure to a field SI variant. To accomplish our goal we used a randomized, blinded, field trial in 2 cohorts of finisher pigs in a multi-site swine production system located in southern Ontario. During the trial, this herd experienced an outbreak of porcine circovirus associated disease (PCVAD). The efficacy of the SI vaccine was assessed through its effect on average daily weight gain, and serological responses to SI over time. The effect of vaccination on pig growth was different in the 2 cohorts. Weight gain was higher in vaccinated pigs than in control pigs in Cohort 1, but was numerically higher for control pigs than for vaccinated pigs in Cohort 2. Vaccination against swine influenza, in a herd experiencing an outbreak of PCVAD, was of questionable value.

The objective of this study was to estimate the apparent and true prevalence of exposure to Toxoplasma gondii in Ontario finisher pigs. During the study period (2001 to 2004), sera from 6048 pigs were tested with a commercial enzyme-linked immunosorbent assay (ELISA); 103 farms were included 1 to 3 times in the study. True prevalence was estimated using a Bayesian approach. Apparent prevalence at the pig level was 1.59% [95% confidence interval (CI): 0.45, 2.99] in 2001, 0.06% (95% CI: 0.00, 0.46) in 2003, and 0.26% (95% CI: 0.00, 0.82) in 2004. Apparent prevalence at the herd-level was 13.7% (95% CI: 7.5, 22.3) in 2001; 1.25% (95% CI: 0.03, 6.77) in 2003, and 3.75% (95% CI: 0.78, 10.6) in 2004. Similarly, posterior Bayesian estimates of true prevalence at the pig level were 1.7% [95% probability interval (PI): 1.2, 2.2] in 2001, 0.2% (95% PI: 0.04, 0.4) in 2003, and 0.3% (95% PI: 0.1, 0.7) in 2004. At the herd level, posterior estimates of prevalence were 11.6% (95% PI: 7.4, 16.8) in 2001, 0% (95% PI: 0.0, 2.5) in 2003, and 1.2% (95% PI: 0.0, 5.0) in 2004 when a herd cut-pointor = 1 was used. Exposure to T. gondii in finishing pig farms in Ontario appears to be infrequent.

SUMMARYThe objectives of this study were to identify factors associated with Salmonella status at the farm, pen, and pig level; explore the nature of variation in the association between the pen-level Salmonella status and pen-level covariates, and pig-level Salmonella status and pig- and pen-level covariates; and to identify the relative importance of factors operating at geographical, farm, and pen level for Salmonella shedding of pigs. For these purposes, samples from 799 pigs and 374 pens on 80 farms in Ontario in 2004 were collected and bacteriologically tested in a cross-sectional study. Census division was the least variable level, and farm the most variable level for shedding. Increased frequency of disinfection and washing with cold water were positively associated with Salmonella positivity, whereas liquid and mash feed and completely closed barns were sparing factors. After farm, pen was the second most variable level for shedding. However, no measured pen-level variables were associated with Salmonella status of pigs or pens. The shedding of Salmonella at the pig level tended to be associated with pig weight, and there was no random variation around this association. Results of this study suggest that a herd test based on bacteriological culture would probably have higher sensitivity if growing animals of lower weight were sampled instead of market weight animals, and this might be beneficial for Salmonella monitoring.

We investigated characteristics of Yersinia enterocolitica infection in Ontario finisher pig herds. Our specific objectives were to estimate or test: prevalence of Y. enterocolitica shedding in finisher pigs, bioserotype distribution, agreement between the herd-level tests based on sampling pig and pooled fecal samples, whether bioserotypes cluster by farms, and whether Y. enterocolitica-positive herds cluster spatially. In total, 3747 fecal samples were collected from 100 farms over the years 2001, 2002, and 2004 (250 total herd visits). Fecal samples were tested by culture and positive isolates were biotyped and serotyped. Apparent pig-level prevalence of Y. enterocolitica was 1.8%, 3.2%, and 12.5% in 2001, 2002, and 2004, respectively. Estimated true pig-level prevalence of Y. enterocolitica was 5.1%, 9.1%, and 35.1% in 2001, 2002, and 2004, respectively. Herd-level prevalence was 16.3%, 17.9%, and 37.5% in 2001, 2002, and 2004, respectively. In all years, the most common bioserotype was 4, O:3, followed by bioserotype 2, O:5,27. Kappa between herd-level status based on pig and pooled samples ranged between 0.51 and 0.68 for biotype 1A and bioserotype 4, O:3, respectively. For 4, O:3, a significant bias in discordant pairs was detected, indicating that pig samples were more sensitive than pooled samples in declaring a herd as positive. Farms tended to be repeatedly positive with the same bioserotype, but positive study farms did not cluster spatially (suggesting lack of between herd transmission and lack of a common geographic risk factor).

We report the methods and findings of a survey of Canadian swine producers summarizing farm-types at-risk of foreign animal disease (FAD) and the routine movement of animals, semen and workers among swine farms, as observed during a 42-day period. Of the 311 producers who returned completed questionnaires, 17% represented swine-herds with no swine or semen movement on or off the farm during the 42 days, 57% were sow herds or farrow-to-finish herds with limited movement onto the farm but movement off the farm, and 26% were swine-herds with movements on and off the farm. A substantial number of premises (>50% in some provinces) with swine also kept other animal species on the same premises. We applied the empirical movement data from the survey in a stochastic simulation model to estimate the number of herds infected and the basic regional distribution of infection that could be expected to occur if the FAD was not detected and routine movements were permitted to occur up to 42 days after infection with a FAD of a single randomly selected herd. Forty-five percent of the simulations did not involve spread beyond the index farm, whereas 34.8% involved spread among five or more farms after 42 days of routine movement.

Three unusual, highly oxygenated novel phenylpropanoids (1−3) and two novel isoflavans, 8-prenylmucronulatol (4) and smiranicin (6), were isolated from Smirnowia iranica together with a previously described isoflavan, glyasperin H (5). The structures were established using homo- and heteronuclear two-dimensional NMR experiments. The isoflavans significantly inhibited the growth of extracellular stages of three Leishmania species in vitro, their activity against the intracellular stages being considerably lower. 8-Prenylmucronulatol (4) showed moderate in vitro toxicity against Plasmodium falciparum, without noticeable erythrocyte membrane effects at the inhibitory concentration. Because of the structural relationship of isoflavans with chalcones and aurones, some of which are potent antiprotozoal agents, the isoflavan skeleton may be a template structure in search for new compounds with leishmanicidal and antiplasmodial activity.

We compared the efficiency of immunophenotyping using flow cytometry (FCM) and a combination of morphologic and immunocytochemical studies for detecting malignant cells in 92 effusions. Cytologic results were as follows: carcinoma cells, 43 specimens; benign, 42 specimens; suggestive of nonepithelial malignancy, 7 specimens. After immunocytochemical analysis, 5 benign specimens were reclassified as malignant and 4 malignant epithelial specimens as benign. With FCM, cells positive for Ber-EP4, B 72.3, AH6, and HB-TN were detected in 28 to 36 (64%-82%) of 44 carcinomas but only 2 to 12 (5%-29%) of 41 benign specimens. Significant association was seen for coexpression. Ber-EP4 and AH6 were the most sensitive; Ber-EP4 was the most specific. The presence of cells positive for 3 of 4 markers strongly suggested malignancy (34/44 carcinoma specimens [77%]; 3/41 reactive specimens [7%]). The presence of cells positive for all 4 markers was diagnostic of malignancy (17/44 malignant specimens [39%]; 0/41 reactive effusions [0%]). FCM and immunocytochemical resultsfor Ber-EP4 expression showed excellent association. FCM is a powerful tool for diagnosing difficult effusions and can quantify coexpression of various markers in fresh specimens. By using established cellular markers coupled with biological markers, FCM also has great promise for experimental purposes.

A randomized double-blind placebo-controlled clinical trial was carried out to determine whether a group of stable children with cerebral palsy (36 males, 21 females; mean age 10 years 11 months, range 5 to 18 years) would improve their motor skills after 12 months of threshold electrical stimulation (TES). Two thirds received active and one third received inactive stimulators. For the primary outcome we constructed a set of plausible motor function tests and studied the change in summary indices of the performance measurements. Tests were videotaped and assessed blindly to record qualitative changes that might not be reflected in performance measurements. We also judged range of motion, degree of spasticity, and muscle growth measured by CT. Fifty seven of 82 outpatients who were able to walk at least with a walker, completed all 12 months of treatment (hemiplegia n=25, diplegia n=32). There was no significant difference between active and placebo treatment in any of the tested groups, nor combined. Visual and subjective assessments favoured TES (ns), whereas objective indices showed the opposite trend. We conclude that TES in these patients did not have any significant clinical effect during the test period.

Lupeol, which shows in vitro inhibitory activity against Plasmodium falciparum 3D7 strain with a 50% inhibitory concentration (IC 50 ) of 27.7 ± 0.5 μM, was shown to cause a transformation of the human erythrocyte shape toward that of stomatocytes. Good correlation between the IC 50 value and the membrane curvature changes caused by lupeol was observed. Preincubation of erythrocytes with lupeol, followed by extensive washing, made the cells unsuitable for parasite growth, suggesting that the compound incorporates into erythrocyte membrane irreversibly. On the other hand, lupeol-treated parasite culture continued to grow well in untreated erythrocytes. Thus, the antiplasmodial activity of lupeol appears to be indirect, being due to stomatocytic transformation of the host cell membrane and not to toxic effects via action on a drug target within the parasite. A number of amphiphiles that cause stomatocyte formation, but not those causing echinocyte formation, were shown to inhibit growth of the parasites, apparently via a mechanism similar to that of lupeol. Since antiplasmodial agents that inhibit parasite growth through erythrocyte membrane modifications must be regarded as unsuitable as leads for development of new antimalarial drugs, care must be exercised in the interpretation of results of screening of plant extracts and natural product libraries by an in vitro Plasmodium toxicity assay.

Cryptotanshinone (1), a quinoid diterpene with a nor-abietane skeleton, and three new natural products, 1beta-hydroxycryptotanshinone (2), 1-oxocryptotanshinone (3), and 1-oxomiltirone (4), were isolated from roots of the Iranian medicinal plant Perovskia abrotanoides. Their structures were established using homo- and heteronuclear two-dimensional NMR experiments, supported by HRMS. The total amount of tanshinones isolated from dry roots of Perovskia abrotanoides was about 1.5%. The compounds exhibited leishmanicidal activity in vitro (IC(50) values in the range 18-47 microM). These findings provide a rationale for traditional use of the roots in Iran as a constituent of poultices for treatment of cutaneous leishmaniasis. The isolated tanshinones also inhibited growth of cultured malaria parasites (3D7 strain of Plasmodium falciparum), drug-sensitive KB-3-1 human carcinoma cell line, multidrug-resistant KB-V1 cell line, and human lymphocytes activated with phytohaemagglutinin A (IC(50) values in the range 5-45 microM). The toxicity of tanshinones toward the drug-sensitive KB-3-1 and the multidrug-resistant KB-V1 cells was the same, indicating that the compounds are not substrates for the P-glycoprotein drug efflux pump.

The objective of the present study was to compare the efficiency of immunophenotyping using flow cytometry (FCM) and immunocytochemistry (ICC) in the detection of macrophages in serous effusions. Cytoblock sections from 90 effusions were stained for the monocyte/macrophage marker CD14, using ICC. Fresh-frozen samples of all cases were analyzed for CD14 expression, using FCM. Epithelial, lymphoid, and mesothelial cell populations were identified using antibodies against Ber-EP4, CD45, and N-cadherin, respectively. Results were compared with clinical parameters and morphological diagnosis. Thirty-nine specimens were cytologically diagnosed as malignant, containing tumor cells of nonhematologic origin, whereas 46 were interpreted as benign. Two additional specimens were diagnosed as indeterminate or suspicious for malignancy, and 3 specimens contained lymphoma cells. CD14-positive cells were detected in 85/90 (94%) of effusions using FCM, and in all 90 specimens using ICC. The percentage of CD14-positive cells was highly variable, but in some specimens was as high as 76% using FCM and 85% using ICC. A good association was observed between the two methods in the detection of CD14-positive cells (P0.001). The presence of macrophages in effusions showed an association with female gender, using both FCM (P = 0.002) and ICC (P = 0.011), but none with effusion site, patient age, clinical and cytological diagnosis, or presence of Ber-EP4-positive cells (P0.05). The presence of Ber-EP4-positive cells showed a strong association with the cytological diagnosis of malignancy (P0.001). In conclusion, macrophages are a significant cell population in effusions, of both benign and malignant etiology, due to both their size and their possible confusion with cancer cells. Both FCM and ICC aid in the recognition of these cells, and thus provide an effective tool for the identification of different cell populations in effusions.

Definitions of epidemiological concepts regarding disease monitoring and surveillance can be found in textbooks on veterinary epidemiology. This paper gives a review of how the concepts: monitoring, surveillance, and disease control strategies are defined. Monitoring and surveillance systems (MO&SS) involve measurements of disease occurrence, and the design of the monitoring determines which types of disease occurrence measures can be applied. However, the knowledge of the performance of diagnostic tests (sensitivity and specificity) is essential to estimate the true occurrence of the disease. The terms, disease control programme (DCP) or disease eradication programme (DEP), are defined, and the steps of DCP/DEP are described to illustrate that they are a process rather than a static MO&SS.

Five indole alkaloids, corynantheidine, corynantheine, dihydrocorynantheine, alpha-yohimbine and corynanthine were isolated from bark of Corynanthe pachyceras K. Schum. (Rubiaceae). The structures were established by spectroscopic methods, including previously unreported assignment of all 1H-NMR resonances by COSY and NOESY experiments. These and related alkaloids showed pronounced activity against Leishmania major promastigotes (IC50 at the micromolar level) but no significant in vitro antiplasmodial activity (against chloroquine-sensitive Plasmodium falciparum). Cytotoxicity assessed with drug sensitive KB-3-1 and multidrug-resistant KB-V1 cell lines was low; the alkaloids are apparently not substrates for the P-glycoprotein (P-170) efflux pump.