A Multiplex RT-PCR Method for the Detection of Reptarenavirus Infection.

Reptarenaviruses cause Boid Inclusion Body Disease (BIBD), a fatal disease of boid snakes with an economic and ecological impact, as it affects both captive and wild constrictor snakes. The clinical picture of BIBD is highly variable but often only limited. Intracytoplasmic inclusion bodies (IB), which develop in most cell types including blood cells, are the pathognomonic hallmark of BIBD; their detection represents the diagnostic gold standard of the disease. However, IBs are not consistently present in clinically healthy reptarenavirus carriers, which can, if undetected, lead to and maintain the spread of the disease within and between snake populations. Sensitive viral detection tools are required for screening and control purposes; however, the genetic diversity of reptarenaviruses hampers the reverse transcription (RT) PCR-based diagnostics. Here, we describe a multiplex RT-PCR approach for the molecular diagnosis of reptarenavirus infection in blood samples. The method allows the detection of a wide range of reptarenaviruses with the detection limit reaching 40 copies per microliter of blood. Using 245 blood samples with a reference RT-PCR result, we show that the technique performs as well as the segment-specific RT-PCRs in our earlier studies. It can identify virus carriers and serve to limit reptarenavirus spreading in captive snake collections.

Artificial intelligence in breast imaging: Current situation and clinical challenges.

Breast cancer ranks among the most prevalent malignant tumours and is the primary contributor to cancer-related deaths in women. Breast imaging is essential for screening, diagnosis, and therapeutic surveillance. With the increasing demand for precision medicine, the heterogeneous nature of breast cancer makes it necessary to deeply mine and rationally utilize the tremendous amount of breast imaging information. With the rapid advancement of computer science, artificial intelligence (AI) has been noted to have great advantages in processing and mining of image information. Therefore, a growing number of scholars have started to focus on and research the utility of AI in breast imaging. Here, an overview of breast imaging databases and recent advances in AI research are provided, the challenges and problems in this field are discussed, and then constructive advice is further provided for ongoing scientific developments from the perspective of the National Natural Science Foundation of China.

A comparison of breast screening performance based on a standardised test, during and outside of the COVID-19 lockdown period.

PURPOSE: In the UK, the COVID-19 pandemicand the resulting lockdown significantly impacted routine breast screening and led to reduced case volumes for breast screening readers. The aim of the study was to evaluate whether breast screening performance on a test-set based assessment scheme (PERFORMS) varied over time, in association with the lockdown period. METHOD: In this retrospective study, performance data were obtained for all breast cancer screening readers in England who completed the PERFORMS schemes based on digital mammography (from 2015 to 2022). Cancer detection (sensitivity), correct return to screen (specificity) and the difference between the sensitivity and specificity were calculated for each breast screener in each PERFORMS scheme. RESULTS: During the study period, 4906 readers participated in eight PERFORMS schemes. Both the cancer detection (H(7) = 775.56, p <.00001) and correct return to screen rate (H(7) = 401.13, p <.00001) varied significantly by scheme. The difference between cancer detection and correct return to screen rate in the scheme administered during the lockdown period was significantly higher than in all other schemes (p <.05, Bonferroni correction applied). CONCLUSIONS: The performance of breast screeners on the PERFORMS test-set based assessment scheme which was administered during the lockdown period, was characterised by markedly high sensitivity and low specificity when compared to their performance on other tests taken pre-COVID and following the lockdown.

Sweet Wine Production from the Side-Stream of Industrial Corinthian Currant Processing: Product Quality, Antioxidant Capacity, and Volatilome.

In the frame of efforts to add value to the Mediterranean currant cultivation and processing sectors, which is essential for their sustainability, sweet wine production is proposed from the finishing side-stream (FSS) of premium quality Corinthian currants, involving complete fermentation using an alcohol-tolerant yeast followed by (i) the addition of FSS to extract sugars or (ii) syrup made from FSS to adjust sweetness. Wine was also made by (iii) ceasing fermentation at the desired sugar level by ethanol addition. The non-fortified wines had 15.2-15.5% ethanol, 115-145 g/L residual sugar, 7.2-7.6 g/L titratable acidity, low volatile acidity (VA; <0.33 g/L), 280-330 mg/L phenolic content (TPC) (as gallic acid), and 23.8-35.6 mg/L antioxidant capacity (AC) (as ascorbic acid). In total, 160 volatiles were identified by SPME GC-MS, including compounds derived from the grapes, the raisin drying, and the fermentation process. The non-fortified wines had better characteristics (mainly VA, AC, and TPC) than the fortified wine, while sweetness adjustment by FSS is the simplest and lowest cost method since it does not involve ethanol or syrup addition. The proposed methods can lead to good quality sweet wines with a characteristic fruity (grape/raisin) flavor that could be commercialized as specialty raisin beverages or liqueurs.

A survey by the European Society of Breast Imaging on radiologists' preferences regarding quality assurance measures of image interpretation in screening and diagnostic mammography.

OBJECTIVES: Quality assurance (QA) of image interpretation plays a key role in screening and diagnostic mammography, maintaining minimum standards and supporting continuous improvement in interpreting images. However, the QA structure across Europe shows considerable variation. The European Society of Breast Imaging (EUSOBI) conducted a survey among the members to collect information on radiologists' preferences regarding QA measures in mammography. MATERIALS AND METHODS: An anonymous online survey consisting of 25 questions was distributed to all EUSOBI members and national breast radiology bodies in Europe. The questions were designed to collect demographic characteristics, information on responders' mammography workload and data about QA measures currently used in their country. Data was analysed using descriptive statistical analysis, the χ2 test, linear regression, and Durbin-Watson statistic test. RESULTS: In total, 251 breast radiologists from 34 countries completed the survey. Most respondents were providing both screening and symptomatic services (137/251, 54.6%), working in an academic hospital (85/251, 33.9%) and reading 1000-4999 cases per year (109/251, 43.4%). More than half of them (133/251, 53%) had established QA measures in their workplace. Although less than one-third (71/251, 28.3%) had to participate in regular performance testing, the vast majority (190/251, 75.7%) agreed that a mandatory test would be helpful to improve their skills. CONCLUSION: QA measures were in place for more than half of the respondents working in screening and diagnostic mammography to evaluate their breast imaging performance. Although there were substantial differences between countries, the importance of having QA in the workplace and implemented was widely acknowledged by radiologists. CLINICAL RELEVANCE STATEMENT: Although several quality assurance (QA) measures of image interpretation are recommended by European bodies or national organisations, the QA in mammography is quite heterogenous between countries and reporting settings, and not always actively implemented across Europe. KEY POINTS: The first survey that presents radiologists' preferences regarding QA measures of image interpretation in mammography. Quality assurance measures in the workplace are better-established for breast screening compared to diagnostic mammography. Radiologists consider that performance tests would help to improve their mammography interpretation skills.

Reptarenavirus S Segment RNA Levels Correlate with the Presence of Inclusion Bodies and the Number of L Segments in Snakes with Reptarenavirus Infection-Lessons Learned from a Large Breeding Colony.

Reptarenaviruses cause boid inclusion body disease (BIBD), a fatal disease particularly impacting captive boa constrictor collections. The development of cytoplasmic inclusion bodies (IBs) comprising reptarenavirus nucleoprotein (NP) in many cell types of affected snakes is characteristic of BIBD. However, snakes can harbor reptarenaviruses without showing IBs, hence representing carriers and a potential source of transmission. The RNA genome of reptarenaviruses comprises a small (S) and a large (L) segment, and the snakes with BIBD commonly carry a swarm of reptarenavirus segments. To design sensitive and reliable tools for the diagnosis of reptarenavirus infection in snake colonies, we used metatranscriptomics to determine the reptarenavirus segments present in a large boa constrictor breeding colony. The analysis identified one reptarenavirus S segment and three L segments in the colony. The sequence data served to design real-time reverse transcription-PCR (RT-PCR) targeting the found S segment. This allowed us to identify all infected animals and to quantify the S segment RNA levels, which we found to correlate with the presence of IBs. We further found a positive correlation between the number of L segments and the S segment RNA level, which could suggest that L segment excess also contributes to the IB formation. Information on cohousing of the snakes showed a clear association of reptarenavirus infection with cohousing in general and cohousing with infected animals. Information on breeding and offspring confirmed that vertical transmission occurred. Furthermore, our data suggest that some animals might be able to clear the infection or at least exhibit transient or intermittent viremia. IMPORTANCE Boid inclusion body disease (BIBD) is caused by reptarenavirus infection, and while reptarenavirus nucleoprotein is the main component of the inclusion bodies (IBs) characteristic of BIBD, not all reptarenavirus-infected snakes demonstrate IBs in their cells. Identification of infected individuals is critical for controlling the spread of the disease; however, the genetic divergence of reptarenaviruses complicates reverse transcription-PCR (RT-PCR)-based diagnostics. Here, we tested a next-generation-sequencing-based approach to establish a tailored "colony-specific" set of diagnostic tools for the detection of reptarenavirus small (S) and large (L) genome segments. With this approach, we could demonstrate that an S-segment-specific RT-PCR is highly effective in identifying the infected individuals. We further found the S segment RNA level to positively correlate with the presence of IBs and the number of L segments, which could direct future studies to identify the BIBD pathogenetic mechanisms.

Air pollution mitigation assessment to inform Cambodia's first clean air plan.

Cambodia's 16.5 million people are exposed to air pollution in excess of World Health Organisation guidelines. The Royal Government of Cambodia has regulated air pollutant emissions and concentrations since 2000, but rapid economic growth and energy consumption means air pollution continues to impact human health. In December 2021, the Ministry of Environment of Cambodia published Cambodia's first Clean Air Plan that outlines actions to reduce air pollutant emissions over the next decade. This work presents the quantitative air pollution mitigation assessment underpinning the identification and evaluation of measures included in Cambodia's Clean Air Plan. Historic emissions of particulate matter (PM2.5, black carbon, organic carbon) and gaseous (nitrogen oxides, volatile organic compounds, sulphur dioxide, ammonia, and carbon monoxide) air pollutants are quantified between 2010 and 2015, and projected to 2030 for a baseline scenario. Mitigation scenarios reflecting implementation of 14 measures included in Cambodia's Clean Air Plan were modelled, to quantify the national reduction in emissions, from which the reduction in ambient PM2.5 exposure and attributable health burdens were estimated. In 2015, the residential, transport, and waste sectors contribute the largest fraction of national total air pollutant emissions. Without emission reduction measures, air pollutant emissions could increase by between 50 and 150% in 2030 compared to 2015 levels, predominantly due to increases in transport emissions. The implementation of the 14 mitigation measures could substantially reduce emissions of all air pollutants, by between 60 and 80% in 2030 compared to the baseline. This reduction in emissions was estimated to avoid approximately 900 (95% C.I.: 530-1200) premature deaths per year in 2030 compared to the baseline scenario. In addition to improving air pollution and public health, Cambodia's Clean Air Plan could also to lead to additional benefits, including a 19% reduction in carbon dioxide emissions, simultaneously contributing to Cambodia's climate change goals.

Performance of Radiologists and Radiographers in Double Reading Mammograms: The UK National Health Service Breast Screening Program.

Background Double reading can be used in screening mammography, but it is labor intensive. There is limited evidence on whether trained radiographers (ie, technologists) may be used to provide double reading. Purpose To compare the performance of radiologists and radiographers double reading screening mammograms, considering reader experience level. Materials and Methods In this retrospective study, performance and experience data were obtained for radiologists and radiographer readers of all screening mammograms in England from April 2015 to March 2016. Cancer detection rate (CDR), recall rate (RR), and positive predictive value (PPV) of recall based on biopsy-proven findings were calculated for first readers. Performance metrics were analyzed according to reader professional group and years of reading experience using the analysis of variance test. P values less than .05 were considered to indicate statistically significant difference. Results During the study period, 401 readers (224 radiologists and 177 radiographers) double read 1 404 395 screening digital mammograms. There was no difference in CDR between radiologist and radiographer readers (mean, 7.84 vs 7.53 per 1000 examinations, respectively; P = .08) and no difference for readers with more than 10 years of experience compared with 5 years or fewer years of experience, regardless of professional group (mean, 7.75 vs 7.71 per 1000 examinations respectively, P = .87). No difference in the mean RR was observed between radiologists and radiographer readers (5.0% vs 5.2%, respectively, P = .63). A lower RR was seen for readers with more than 10 years of experience compared with 5 years or fewer, regardless of professional group (mean, 4.8% vs 5.8%, respectively; P = .001). No variation in PPV was observed between them (P = .42), with PPV values of 17.1% for radiologists versus 16.1% for radiographers. A higher PPV was seen for readers with more than 10 years of experience compared with 5 years or less, regardless of professional group (mean, 17.5% and 14.9%, respectively; P = .02). Conclusion No difference in performance was observed between radiographers and radiologists reading screening mammograms in a program that used double reading. Published under a CC BY 4.0 license Online supplemental material is available for this article. See also the editorial by Hooley and Durand in this issue.

Integrated climate change and air pollution mitigation assessment for Togo.

Togo, in west Africa, is vulnerable to the impacts of climate change, but has made a negligible contribution to causing it. Togo ratified the Paris Agreement in 2017, committing to submit Nationally Determined Contributions (NDCs) that outline Togo's climate change mitigation commitment. Togo's capital, Lomé, as well as other areas of Togo have ambient air pollutant levels exceeding World Health Organisation guidelines for human health protection, and 91 % of Togolese households cook using solid biomass, elevating household air pollution exposure. In Togo's updated NDC, submitted in 2021, Togo acknowledges the importance and opportunity of achieving international climate change mitigation targets in ways that improve air quality and achieve health benefits for Togo's citizens. The aim of this work is to evaluate priority mitigation measures in an integrated assessment of air pollutant, Short-Lived Climate Pollutant (SLCP) and Greenhouse Gas (GHG) emissions to identify their effectiveness in simultaneously reducing air pollution and Togo's contribution to climate change. The mitigation assessment quantifies emissions for Togo and Grand Lomé from all major source sectors for historical years between 2010 and 2018, for a baseline projection to 2030 and for mitigation scenarios evaluating ten mitigation measures. The assessment estimates that Togo emitted ~21 million tonnes of GHG emissions in 2018, predominantly from the energy and Agriculture, Forestry and Other Land Use sectors. GHG emissions are projected to increase 42 % to 30 million tonnes in 2030 without implementation of mitigation policies and measures. The implementation of the ten identified priority mitigation measures could reduce GHG emissions by ~20 % in 2030 compared to the baseline, while SLCPs and air pollutants were estimated to be reduced more, with a more than 75 % reduction in black carbon emissions in 2030. This work therefore provides a clear pathway by which Togo can reduce its already small contribution to climate change while simultaneously achieving local benefits for air quality and human health in Togo and Grand Lomé.

Translation and validation of an ageism scale for dental students in Switzerland.

PURPOSE: This study aimed to validate an ageism scale for dental students in Switzerland. METHODS: The original scale was first translated to German and then evaluated by 14 experts for relevance; four items were eliminated. Validation of the resulting questionnaire was performed at three Swiss dental schools. Principal component analysis (PCA) was performed; Cronbach's alpha (α) was used to assess the internal consistency reliability, and Pearson's coefficient to identify any correlations with demographic parameters (P < 0.05). RESULTS: PCA revealed 11 items among 4 factors (Overall: 11 items, α = 0.63, variance = 56.6%; Factor #1 (F1): items = 3, α = 0.64; Factor #2 (F2): items = 3, α = 0.42; Factor #3 (F3): items = 3, α = 0.35; Factor #4 (F4): items = 2, α = 0.37). F1 and F4 were correlated with clinical experience (F1: P = 0.042; F4: P = 0.006) and participation in a gerodontology course (F1: P = 0.021; F4: P = 0.004). F1 was correlated with experience of dealing with the elderly (P = 0.031), while residence locality was correlated with F3 (P = 0.047) and F4 (P = 0.043). F2 was correlated with the presence of elderly in the family (P = 0.047). CONCLUSION: The translated dental ageism questionnaire for Switzerland resulted in an 11-item scale with acceptable reliability.

Gammaherpesvirus Infections in Cattle in Europe.

The genus Macavirus, subfamily Gammaherpesvirinae, comprises ungulate viruses that infect domestic and wild ruminants and swine. They cause asymptomatic latent infections in reservoir hosts and malignant catarrhal fever in susceptible species. Lung, spleen, bronchial lymph node, and tongue were collected from 448 cattle (348 necropsied, 100 slaughtered) in Switzerland, United Kingdom, Finland, Belgium, and Germany to determine their infection with bovine herpesvirus-6 (BoHV-6) and gammaherpesviruses of other ruminants, i.e., ovine herpesvirus-1 and -2, caprine herpesvirus-2, and bison lymphotropic herpesvirus, using quantitative PCR. Only BoHV-6 was detected, with an overall frequency of 32%, ranging between 22% and 42% in the different countries. Infection was detected across all ages, from one day after birth, and was positively correlated with age. There was no evidence of an association with specific disease processes. In positive animals, BoHV-6 was detected in all organs with high frequency, consistently in the lungs or spleen. Viral loads varied substantially. In BoHV-6-positive gravid cows, organs of fetuses tested negative for infection, indicating that the virus is not vertically transmitted. Our results confirm previous data indicating that BoHV-6 is a commensal of domestic cattle not associated with disease processes and confirm that infections with other macaviruses are rare and sporadic.

The relationship between missed breast cancers on mammography in a test-set based assessment scheme and real-life performance in a National Breast Screening Programme.

PURPOSE: This retrospective study determined whether a test-set based assessment scheme (PERFORMS) used in a national breast screening programme could be used to predict real-life performance by investigating if the number of cancers missed by mammography readers in real-life related to the number of cancers missed in the PERFORMS test-set and whether real-life reading volumes affected performance. METHOD: Data was obtained from consenting readers in the screening programme in England (NHSBSP) where double reading is standard. The rate of cancers missed by individual first readers but correctly identified by second readers was compared with the number of cancers missed in the PERFORMS test-set over a 3-year period. NHSBSP readers are required to interpret at least 1500 cases per year as a first reader, so results were compared between readers who exceeded this target and those that did not. Parametric and non-parametric correlations were calculated. RESULTS: Amongst the 536 readers, there was a highly significant positive correlation between the real-life and PERFORMS test-set missed cancer metrics (Pearson Correlation = 0.228, n = 536, p < .0001, Spearman's rho = 0.265, n = 536, p < .0001). There was no significant difference in rates of missed cancers between the 452 readers who exceeded the 1500 first read per year target and those who did not (t(94.2) = -1.87, p = .0643, r = 0.19). CONCLUSIONS: The use of a test-set based assessment scheme accurately reflects real-life mammography reading performance, indicating that it can be a useful tool in identifying poor reader performance.

Beef abattoir interventions in a risk-based meat safety assurance system.

In risk-based meat safety assurance system, the use of interventions is intended to accomplish the meat safety targets on chilled carcasses, particularly in situations when an abattoir is unable to sufficiently reduce risks arising from specific farms/animal batches by using process hygiene alone. Furthermore, interventions are considered whenever food safety authorities identify meat production processes associated with high risks for consumers. This paper overviews the role of beef interventions in a risk-based, meat safety assurance system. Cattle hide interventions (chemical hide washes and microbial immobilisation treatment with shellac) and beef carcass interventions (pasteurisation treatments with hot water and/or steam and organic (lactic) acid washes), show consistent reduction effects of aerobic bacteria and faecal indicators and reduced prevalences of naturally present VTEC and Salmonella. The review also identified interventions where there was a lack of data and further research was needed, and other contextual factors to inform the risk management decisions for further development of risk-based meat safety assurance system.

Colony Stimulating Factors in Early Feline Infectious Peritonitis Virus Infection of Monocytes and in End Stage Feline Infectious Peritonitis; A Combined In Vivo And In Vitro Approach.

Feline coronavirus (FCoV) infection initiates monocyte-associated viremia and viral persistence. Virus-infected, -activated monocytes also trigger feline infectious peritonitis (FIP), a fatal systemic disease of felids typified by granulomatous (peri)phlebitis. Currently, the exact mechanisms inducing monocyte activation and FIP are unknown. This study attempted to identify the potential immediate effect of virulent FCoV on colony-stimulating factor (CSF) (granulocyte (G)-CSF, monocyte (M)-CSF and granulocyte-monocyte (GM)-CSF levels through in vitro assessment, alongside prototypical pro- and anti-inflammatory mediators (interleukin (IL)-1, IL-6, IL-12p40, tumor necrosis factor (TNF)-α, and IL-10); this was assessed alongside the in vivo situation in the hemolymphatic tissues of cats euthanized with natural end-stage FIP. For the in vitro work, isolated monocytes from SPF cats were cultured short-term and infected with the FIP virus (FIPV) strain DF2. Mediator transcription was assessed by quantitative reverse transcriptase PCR (RT-qPCR) at 3, 6 and 9 h post infection (hpi), and in the post-mortem samples of bone marrow, spleen, and mesenteric lymph nodes (MLN) of cats with FIP. We observed limited and transient changes in cytokine transcription in monocytes after infection, i.e., a significant increase of IL-6 at 3 hpi and of GM-CSF over the 3 and 6 hpi period, whereas M-CSF was significantly decreased at 9 hpi, with a limited effect of age. The findings indicate that the infection induces expansion of the monocyte/macrophage population, which would ensure the sufficient supply of cells for consistent viral replication. In natural disease, the only upregulation was of G-CSF in the MLN, suggesting either immune exhaustion or an active downregulation by the host as part of its viral response.

Antibody response in snakes with boid inclusion body disease.

Boid Inclusion Body Disease (BIBD) is a potentially fatal disease reported in captive boid snakes worldwide that is caused by reptarenavirus infection. Although the detection of intracytoplasmic inclusion bodies (IB) in blood cells serves as the gold standard for the ante mortem diagnosis of BIBD, the mechanisms underlying IB formation and the pathogenesis of BIBD are unknown. Knowledge on the reptile immune system is sparse compared to the mammalian counterpart, and in particular the response towards reptarenavirus infection is practically unknown. Herein, we investigated a breeding collection of 70 Boa constrictor snakes for BIBD, reptarenavirus viraemia, anti-reptarenavirus IgM and IgY antibodies, and population parameters. Using NGS and RT-PCR on pooled blood samples of snakes with and without BIBD, we could identify three different reptarenavirus S segments in the collection. The examination of individual samples by RT-PCR indicated that the presence of University of Giessen virus (UGV)-like S segment strongly correlates with IB formation. We could also demonstrate a negative correlation between BIBD and the presence of anti-UGV NP IgY antibodies. Further evidence of an association between antibody response and BIBD is the finding that the level of anti-reptarenavirus antibodies measured by ELISA was lower in snakes with BIBD. Furthermore, female snakes had a significantly lower body weight when they had BIBD. Taken together our findings suggest that the detection of the UGV-/S6-like S segment and the presence of anti-reptarenavirus IgY antibodies might serve as a prognostic tool for predicting the development of BIBD.

Reliability of indicators of sheep welfare assessed by a group observation method.

Assessments of animal-based outcomes form the core of routine veterinary clinical examinations and are being increasingly used as indicators of animal welfare. A method of group observation that did not require gathering and handling of individual sheep, was used to assess eight animal-based indicators of sheep welfare: demeanour, skin irritation, wool loss, excessive panting, coughing, lameness, and cleanliness of the ventral abdominal and 'breech' (perineum/gluteal/caudal hindlimb) areas. The inter-observer reliability of two or three observers who independently assessed these indicators was tested on 2406 adult sheep and growing lambs across 36 farms and the intra-observer reliability of an experienced, veterinary assessor--the 'test standard observer'--was assessed on 88 adult sheep during four on-farm assessments. Observer reliability was evaluated using Cronbach's alpha (α) comparison of the recorded proportion (%) of sheep affected by each welfare condition and by binomial logistic regression modelling. High levels of inter-observer reliability were identified for the assessment of group lameness (α 0.76-1.00) and cleanliness of the breech area (α 0.97-1.00). Excellent intra-observer reliability was determined for lameness (α 0.99), cleanliness of the breech area (α 0.97), demeanour (α 1.00) and wool loss (α 1.00). In addition, proportion data and logistic regression models identified few between-observer differences. The results suggest that welfare outcomes based on observations of the behaviour and physical appearance of individual animals within a group may offer a reliable and feasible measurement tool for the on-farm assessment of sheep welfare.