Comparison of three mathematical models to estimate lactation performance in dairy cows.

Milk yield dynamics and production performance reflect how dairy cows cope with their environment. To optimize farm management, time-series of individual cow milk yield have been studied in the context of precision livestock farming, and many mathematical models have been proposed to translate raw data into useful information for the stakeholders of the dairy chain. To gain better insights on the topic, this study aimed at comparing 3 recent methods that allow to estimate individual cow potential lactation performance, using daily data recorded by the automatic milking systems of 14 dairy farms (7 Holstein, 7 Italian Simmental) from Belgium, the Netherlands, and Italy. An iterative Wood model (IW), a perturbed lactation model (PLM), and a quantile regression (QR) were compared in terms of estimated total unperturbed (i.e., expected) milk production and estimated total milk loss (relative to unperturbed yield). The IW and PLM can also be used to identify perturbations of the lactation curve and were thus compared in this regard. The outcome of this study may help a given end-user in choosing the most appropriate method according to their specific requirements. If there is a specific interest in the post-peak lactation phase, IW can be the best option. If one wants to accurately describe the perturbations of the lactation curve, PLM can be the most suitable method. If there is need for a fast and easy approach on a very large data set, QR can be the choice. Finally, as an example of application, PLM was used to analyze the effect of cow parity, calving season, and breed on their estimated lactation performance.

Sensor-based behavioral patterns can identify heat-sensitive lactating dairy cows.

Heat stress impairs the health and performance of dairy cows, yet only a few studies have investigated the diversity of cattle behavioral responses to heat waves. This research was conducted on an Italian Holstein dairy farm equipped with precision livestock farming sensors to assess potential different behavioral patterns of the animals. Three heat waves, defined as at least five consecutive days with mean daily temperature-humidity index higher than 72, were recorded in the farm area during the summer of 2021. Individual daily milk yield data of 102 cows were used to identify "heat-sensitive" animals, meaning the cows that, under a given heat wave, experienced a milk yield drop that was not linked with other health events (e.g., mastitis). Milk yield drops were detected as perturbations of the lactation curve estimated by iteratively using Wood's equation. Individual daily minutes of lying, chewing, and activity were retrieved from ear-tag-based accelerometer sensors. Semi-parametric generalized estimating equations models were used to assess behavioral deviations of heat-sensitive cows from the herd means under heat stress conditions. Heat waves were associated with an overall increase in the herd's chewing and activity times, along with an overall decrease of lying time. Heat-sensitive cows spent approximately 15 min/days more chewing and performing activities (p < 0.05). The findings of this research suggest that the information provided by high-frequency sensor data could assist farmers in identifying cows for which personalized interventions to alleviate heat stress are needed.

Relationship between sex and clinical and imaging features of early axial spondyloarthritis: results from a 48 month follow-up (Italian arm of the SPondyloArthritis Caught Early (SPACE) study).

OBJECTIVES: To assess the association between sex and clinical and disease activity indices, and X-rays and magnetic resonance imaging (MRI) features, in early-stage axial spondyloarthritis (axSpA). METHOD: Baseline data analysis was conducted on the Italian SPACE cohort, including patients with chronic back pain (duration ≥ 3 months and ≤ 2 years; onset < 45 years). Patients underwent MRI and X-rays of the sacroiliac joints (SIJs) to establish the diagnosis of axSpA, according to Assessment of SpondyloArthritis international Society criteria and physician's judgement. Clinical features, disease activity and functional indices, and images were collected at baseline and yearly during 48 months. Spinal and SIJ X-rays and MRI images were scored by two readers following Spondyloarthritis Research Consortium of Canada (SPARCC), modified Stoke Ankylosing Spondylitis Spinal Score, and modified New York criteria. Characteristics of axSpA patients according to sex (male/female) were compared over time using descriptive statistics. RESULTS: Ninety-one patients had axSpA (83.5% non-radiographic; 16.5% radiographic); 47.3% were male. Males were younger, with shorter duration of axial symptoms, and more frequently had HLA-B27 positivity, radiographic sacroiliitis with a bilateral/symmetric pattern, and more signs of spondylitis. Females more frequently showed peripheral/entheseal involvement and the non-radiographic phenotype. Males showed increased pelvic/spinal radiographic progression and more often had active sacroiliitis on MRI. Although the frequency of inflammatory corner lesions did not differ between males and females, localization varied, with more cervical/thoracic MRI-spine lesions in females and more lumbar lesions in males. We observed a significant downward trend of SPARCC SIJ/spine scores in all patients, irrespective of sex. More fat lesions were observed on MRI-spine in females and on MRI-SIJ in males. CONCLUSION: Sex was associated with distinct axSpA features: females showed low-grade radiographic sacroiliitis and spinal progression, and a higher prevalence of cervical and thoracic spine MRI signs.

A novel nomogram predicting lymph node invasion among patients with prostate cancer: The importance of extracapsular extension at multiparametric magnetic resonance imaging.

PURPOSE: To develop a novel risk tool that allows the prediction of lymph node invasion (LNI) among patients with prostate cancer (PCa) treated with robot-assisted radical prostatectomy (RARP) and extended pelvic lymph node dissection (ePLND). METHODS: We retrospectively identified 742 patients treated with RARP + ePLND at a single center between 2012 and 2018. All patients underwent multiparametric magnetic resonance imaging (mpMRI) and were diagnosed with targeted biopsies. First, the nomogram published by Briganti et al. was validated in our cohort. Second, three novel multivariable logistic regression models predicting LNI were developed: (1) a complete model fitted with PSA, ISUP grade groups, percentage of positive cores (PCP), extracapsular extension (ECE), and Prostate Imaging Reporting and Data System (PI-RADS) score; (2) a simplified model where ECE score was not included (model 1); and (3) a simplified model where PI-RADS score was not included (model 2). The predictive accuracy of the models was assessed with the receiver operating characteristic-derived area under the curve (AUC). Calibration plots and decision curve analyses were used. RESULTS: Overall, 149 patients (20%) had LNI. In multivariable logistic regression models, PSA (OR: 1.03; P= 0.001), ISUP grade groups (OR: 1.33; P= 0.001), PCP (OR: 1.01; P= 0.01), and ECE score (ECE 4 vs. 3 OR: 2.99; ECE 5 vs. 3 OR: 6.97; P< 0.001) were associated with higher rates of LNI. The AUC of the Briganti et al. model was 74%. Conversely, the AUC of model 1 vs. model 2 vs. complete model was, respectively, 78% vs. 81% vs. 81%. Simplified model 1 (ECE score only) was then chosen as the best performing model. A nomogram to calculate the individual probability of LNI, based on model 1 was created. Setting our cut-off at 5% we missed only 2.6% of LNI patients. CONCLUSIONS: We developed a novel nomogram that combines PSA, ISUP grade groups, PCP, and mpMRI-derived ECE score to predict the probability of LNI at final pathology in RARP candidates. The application of a nomogram derived cut-off of 5% allows to avoid a consistent number of ePLND procedures, missing only 2.6% of LNI patients. External validation of our model is needed.

Topics and trends in Mountain Livestock Farming research: a text mining approach.

Pasture-based and small-scale livestock farming systems are the main source of livelihood in the mountain primary sector, ensuring socioeconomic sustainability and biodiversity in rural communities throughout Europe and beyond. Mountain livestock farming (MLF) has attracted substantial research efforts from a wide variety of scientific communities worldwide. In this study, the use of text mining and topic modelling analysis drew a detailed picture of the main research topics dealing with MLF and their trends over the last four decades. The final data corpus used for the analysis counted 2 679 documents, of which 92% were peer-reviewed scientific publications. The number of scientific outputs in MLF doubled every 10 years since 1980. Text mining found that milk, goat and sheep were the terms with the highest weighed frequency in the data corpus. Ten meaningful topics were identified by topic analysis: T1-Livestock management and vegetation dynamics; T2-Animal health and epidemiology; T3-Methodological studies on cattle; T4-Production system and sustainability; T5-Methodological studies; T6-Wildlife and conservation studies; T7-Reproduction and performance; T8-Dairy/meat production and quality; T9-Land use and its change and T10-Genetic/genomic studies. A hierarchical clustering analysis was performed to explore the interrelationships among topics, and three main clusters were identified: the first focused on sustainability, conservation and socioeconomic aspects (T4; T6 and T9), the second was related to food production and quality (T7 and T8) and the last one considered methodological studies on mountain flora and fauna (T1; T2; T3; T5 and T10). The 10 topics identified represent a useful and a starting source of information for further and more detailed analysis (e.g. systematic review) of specific research or geographical areas. A truly holistic and interdisciplinary research approach is needed to identify drivers of change and to understand current and future challenges faced by livestock farming in mountain areas.

A survey on sensor systems used in Italian dairy farms and comparison between performances of similar herds equipped or not equipped with sensors.

Sensor systems (SS) were developed over the last few decades to help dairy farmers manage their herds. Such systems can provide both data and alerts to several productive, behavioral, and physiological indicators on individual cows. Currently, there is still a lack of knowledge on both the proportion of dairy farms that invested in SS and type of SS installed. Additionally, it is still unclear whether the performances of herds equipped with SS differ from those of similar herds managed without any technological aid. Therefore, the aims of this study were (1) to provide an insight into SS spread among Italian dairy farms and (2) to analyze the performances of similar herds equipped or not equipped with SS. To reach the former goal, a large survey was carried out on 964 dairy farms in the northeast of Italy. Farmers were interviewed by the technicians of the regional breeders association to collect information on the type of SS installed on farms and the main parameters recorded. Overall, 42% of the surveyed farms had at least 1 SS, and most of them (72%) reared more than 50 cows. Sensors for measuring individual cow milk yield were the most prevalent type installed (39% of the surveyed farms), whereas only 15% of farms had SS for estrus detection. More sophisticated parameters, such as rumination, were automatically monitored in less than 5% of the farms. To reach the latter goal of the study, a subset of 100 Holstein dairy farms with similar characteristics was selected: half of them were equipped with SS for monitoring at least individual milk yield and estrus, and the other half were managed without any SS. Average herd productive and reproductive data from official test days over 3 yr were analyzed. The outcomes of the comparison showed that farms with SS had higher mature-equivalent milk production. Further clustering analysis of the same 100 farms partitioned them into 3 clusters based on herd productive and reproductive data. Results of the Chi-squared test showed that the proportion of farms equipped with SS was greater in the cluster with the best performance (e.g., higher milk yield and shorter calving interval). However, the presence of a few farms equipped with SS in the least productive cluster for the same parameters pointed out that although the installation of SS may support farmers in time- and labor-saving or in data recording, it is not a guarantee of better herd performance.

An insight into the dairy chain of a Protected Designation of Origin cheese: The case study of Asiago cheese.

The Protected Designation of Origin (PDO) label of the European Union safeguards and guarantees top-quality traditional agri-food products that must be manufactured in a specific region according to traditional production methods. Production specifications of PDO cheeses are often focused on the cheese-making process and lack information on the dairy farming system that is upstream of the chain. This case study aimed to analyze and cluster the dairy farms that supply milk to the chain of Asiago, an internationally known PDO cheese of northeastern Italy. A large survey involving all of the cheese factories of the Asiago PDO chain was made in 2017. Each cheese factory submitted a questionnaire to its supplying dairy farmers concerning (1) farm facilities and herd management and (2) feeding program of lactating cows. Results from 517 farms were processed; there were 67 ± 27% (mean ± standard deviation) respondents per cheese factory. Four clusters of dairy farms were identified by hierarchical clustering analysis. Cluster 1 (8% of the surveyed farms) and cluster 2 (22%) are small in size and low in yield, representing the traditional milk production system; farms are mainly located on mountains or hills and have autochthonous dual-purpose breeds mostly housed in tiestall barns. By rearing cattle of endangered breeds and feeding cows primarily with forages produced on-farm together with the use of pasture, these clusters, and especially cluster 1, have shown to provide essential ecosystem services for landscape and biodiversity preservation in the alpine areas. Clusters 3 and 4 (34 and 36% of the surveyed farms, respectively) gather medium-scale farms mainly located in the lowland that operate according to modern management and housing systems and rear high-producing dairy cows. These cows are mainly fed total mixed rations based on corn silage, but the dietary forage:concentrate ratio is kept relatively high, as farmers are more interested in producing high-quality milk for cheese-making than pushing for yield. Regardless of the cluster allocation, a considerable cow longevity, which is a recognized "iceberg indicator" of cattle well-being, was highlighted. This study showed that different farming systems may lay behind a single PDO cheese. The knowledge of their characteristics is important to reinforce the PDO production specifications as well as to distinguish and protect niche products that come from specific groups of farms that provide essential ecosystem services.

SARS-CoV-2 infection in patients with autoimmune rheumatic diseases in northeast Italy: A cross-sectional study on 916 patients.

BACKGROUND: Whether patients with autoimmune rheumatic diseases (ARD) have a higher risk for SARS-CoV-2 infection (COVID-19) and how SARS-CoV-2 pandemic impacts on adherence to therapy has not been fully elucidated. We assessed the rate and clinical presentation of COVID-19, and adherence to therapy in a large cohort of patients with ARD followed-up in a tertiary University-Hospital in Northeast Italy. METHODS: Between April 9th and April 25th, 2020, after SARS-CoV-2 infection peak, a telephone survey investigating the impact of COVID-19 on patients with systemic lupus erythematosus (SLE), systemic sclerosis (SSc), rheumatoid arthritis (RA), ANCA-associated vasculitis (AAV), and idiopathic inflammatory myopathies (IIM) was administered. Demographics, disease activity status, therapy, occupational exposure, and adherence to social distancing advise were also collected. RESULTS: 916 patients (397 SLE, 182 AAV, 176 SSc, 111 RA, 50 IIM) completed the survey. 148 patients developed at least one symptom compatible with COVID-19 (cough 96, sore throat 64, fever 64, arthromyalgias 59, diarrhea 26, conjunctivitis 18, ageusia/hyposmia, 18). Among the 916 patients, 65 (7.1%) underwent SARS-CoV-2 nasopharyngeal swab (18 symptomatic and 47 asymptomatic), 2 (0.21%) tested positive, a proportion similar to that observed in the general population of the Veneto region. No deaths occurred. 31 patients (3.4%) withdrew ≥1 medication, mainly immunosuppressants or biologics. Adoption of social distancing was observed by 860 patients (93.9%), including 335 (36.6%) who adopted it before official lockdown. CONCLUSIONS: COVID-19 incidence seems to be similar in our cohort compared to the general population. Adherence to therapy and to social distancing advise was high.

Health, behaviour and growth performance of Charolais and Limousin bulls fattened on different types of flooring.

Intensive fattening of late-maturing breeds on concrete or rubberized slatted floors is the prevalent beef production system in mainland Europe. The rationale behind this study is that specific beef breeds with different slaughter weights might have a diverse response to different flooring systems. The study aimed at assessing whether growth performance, health, behaviour and claw condition of two beef breeds, Charolais (CH) and Limousine (LIM), would be affected by their housing on concrete (CS) or rubber-covered (RCS) fully slatted floor. A total of 228 CH (116 on CS; 112 on RCS) and 115 LIM (57 on CS; 58 on RCS) were housed in four and two commercial farms, respectively, in groups of 9.0 ± 2.1 animals/pen with an average space allowance of 3.1 ± 0.2 m2. Draining gaps of CS and RCS pens were 16.9 ± 1.7% and 11.6 ± 1.2% of the total surface, respectively. Bulls of both breeds had similar initial body weight (429.4 ± 31.5 kg for CH; 369.6 ± 31.7 kg for LIM), and they were slaughtered when they reached suitable finishing. Charolais had a higher final body weight (BW) than LIM (750.8 ± 8.6 v. 613.7 ± 10.9 kg; P < 0.01), and bulls of both breeds on RCS had higher average daily gain than on CS (1.47 ± 0.02 v. 1.39 ± 0.02 kg/day; P < 0.05). The percentage of bulls early culled or treated for locomotor disorders were reduced by RCS only for LIM, while RCS tended to prevent the occurrence of bursitis for both breeds. During two 8-h behavioural observations, bulls on RCS performed more head butt/displacements and chases than on CS, and they reduced the frequency of abnormal lying down events. The use of RCS increased mounts' frequency only in LIM, while its reduced drainage capacity impaired only the cleanliness of CH. Postmortem hoof inspection showed longer claw dorsal wall and diagonal lengths, and sharper toe angles for CH on RCS than LIM on both floors. Results of this study point out that fully slatted floors, regardless of being rubberized or not, are not suitable for bulls finished at a final BW above 700 kg due to their detrimental effects on health and welfare. The use of RCS could be recommended as an alternative to CS only if bulls are slaughtered at a lower final BW (around 600 kg), like in the case of LIM breed.

Risk factors associated with beef cattle losses on intensive fattening farms in Austria, Germany and Italy.

The aim of this study was to investigate management and feeding practices associated with on-farm loss rate (mortality) on 63 beef cattle farms in Austria, Germany and Italy with housing systems other than fully slatted pens. Information on mortality and 56 categorised factors relating to the cleanliness of animal facilities, health and feeding management, animal-human interaction, cattle transport and origin were gathered during on-farm visits. Samples of total mixed rations (TMRs) were collected and analysed for chemical composition and particle size distribution. Twenty-eight categorised factors were removed from the initial 56 due to exclusion criteria (missing data≥20% and/or monolevel factors with≥80% answers in one category). Mortality was the response variable in the risk factor analysis and the remaining 10 continuous covariates from TMR analyses and 28 categorised factors were independent predictors. Mean (±standard deviation) mortality, representing the proportion of dead, euthanased and early culled animals over the total number of animals bought in or reared in the previous year, was 2.8±3.5%. Fourteen factors were significantly associated with mortality in the bivariable analyses; seven factors were not considered further in the multivariable analysis due to collinearity. None of the factors related to TMR were associated with mortality. Four categorical factors, referring to biosecurity measures and management, were retained in the final multivariable model, with country effect. Buying cattle from only one farm, no mixing of animals during transport, presence of a dedicated sick pen and keeping production records were associated with lower percentage mortality.

A survey of selected animal-based measures of dairy cattle welfare in the Eastern Alps: Toward context-based thresholds.

In the Alps, traditional dairy farms are small-scale operations where vertical transhumance from valley indoor housing systems to highland pasture-based systems is still practiced in summer. Vertical transhumance implies a substantial change of environment, available resources, and management practices from one season to another. In such systems, animal-based welfare measures need to be monitored throughout the year to capture the variation of welfare outcomes, based on which targeted welfare management plans can be implemented. Because the Welfare Quality assessment approach has been tailored to indoor housing and intensive farming systems, the European Food Safety Authority recently developed a welfare assessment protocol for small-scale dairy cattle farms adapted after the Welfare Quality framework. The aim of this study was to assess nonbehavioral animal-based measures as defined by this protocol at different time points for transhumant systems in the Alps. In total, 18 animal-based measures were assessed before, during, and after the mountain pasture period in a sample of 67 small-scale dairy cattle farms practicing vertical transhumance in neighboring provinces of Austria, Italy, and Slovenia. Significant differences between assessments were identified for dirtiness of legs and teats, hairless patches, lesions and swellings, claw condition, ocular discharge, and diarrhea whereas BCS, lameness/severe lameness, vulvar discharge, nasal discharge, and hampered respiration were unchanged between seasons. In addition, a benchmarking exercise was carried out to identify relative boundaries (worst quartile thresholds) for each animal-based measure and to contribute to the discussion about achievable welfare outcomes for the 2 husbandry conditions that characterize a transhumant system. Worst quartile thresholds indicated a high prevalence of dirtiness (>80%) when cows were kept indoors, high prevalence of hairless patches (65%) before pasture turnout, and high prevalence of very lean cows (>13%) throughout the assessments. On the other hand, the best quartile thresholds for most clinical conditions suggested that high welfare standards (zero prevalence) are widely achievable in mountain farms practicing vertical transhumance during all assessments. The thresholds identified through benchmarking should serve as the basis for an effective context-based welfare management strategy promoting continuous welfare improvement on-farm.

A comparative evaluation of a new fully automated assay for von Willebrand factor collagen binding activity to an established method.

INTRODUCTION: Laboratory diagnosis of von Willebrand disease (VWD) is made by the measurement of von Willebrand factor (VWF) protein level and its activities. Current VWF activity tests include ristocetin cofactor and collagen binding (VWF:CB) assays. AIM: We have undertaken an evaluation of a new fully automated VWF:CB assay relative to an established enzyme-linked immunosorbent assay (ELISA) method. METHODS: The two analytical systems operate with different detection principles: a chemiluminescent method performed on ACL AcuStar Analyzer (the former) and a colorimetric ELISA by Asserachrom Stago (the latter) (type III collagen from human placenta). The HemosIL AcuStar VWF:CB assay is a chemiluminescent 2-step immunoassay that uses magnetic particles coated with a type III collagen triple-helical peptide. VWF:CB levels were determined in 50 healthy subjects and 100 VWD patients (22 type 1, 73 type 2 and 5 type 3). RESULTS: Eleven VWD samples reported VWF:CB values below the lower detection limit of one or both methods. The new method showed a good correlation with the ELISA method (r > .9, mean bias 3.85 IU/dL) in both healthy and VWD samples. One of 150 samples gave inconsistent results using the two assays, leading to an uncertain diagnosis of VWD type 1 (ELISA method) or type 2 MCB (fully automated method). CONCLUSION: The new assay is rapid and simple to use, with its ready-to-use reagent cartridges. This VWF:CB assay, in addition to the measurement of VWF:Ag and VWF:RCo made on the same platform, gives additional information for the diagnosis of VWD in both nonspecialized and reference laboratories.

Validation of the Italian version of the Non-Communicating Children's Pain Checklist-Postoperative Version.

BACKGROUND: This study evaluated the validity and reliability of the Italian version of the Non-Communicating Children's Pain Checklist-Postoperative version (I-NCCPC-PV). METHODS: The original NCCPC-PV version was translated into Italian following the guidelines for "the translation, adaptation, and validation of instruments or scales for cross-cultural healthcare research". We tested the Italian NCCPC-PV version (I-NCCPC-PV) in 40 children (3-18 years of age) with severe to profound Intellectual Disability and no verbal communication. Each child's behavior was observed by a parent or caregiver and by an external observer in a quiet situation and a painful one. They independently assessed the child's level of pain using the translated Italian version of the NCCPCPV (I-NCCPC-PV). RESULTS: The results from 80 assessments showed that children's behavioral signs differed significantly between painful and calm situations (p < 0.001). The inter-rater reliability was poor in a quiet condition (ICC 0.62) and fair in a painful situation (ICC 0.77). The inter-rater agreement was good in both calm and painful conditions (72.50% and 77.50% respectively). CONCLUSION: The Italian version of the NCCPC-PV (I-NCCPC-PV) can be used for pain assessment in children with Intellectual Disability who lack verbal communication.

Effect of a ceiling fan ventilation system on finishing young bulls' health, behaviour and growth performance.

This research aimed at assessing the effects of a ceiling fan ventilation system on health, feeding, social behaviour and growth response of finishing young bulls fattened indoors during a mild summer season. A total of 69 Charolais young bulls were housed in six pens without any mechanical ventilation system (Control) and in six pens equipped with ceiling fans. The experimental period lasted 98 days from June until mid-September 2014. Four experimental days were considered in order to assess the effect of the ventilation system under two different microclimatic conditions: 2 alert days at monthly interval with temperature humidity index (THI) between 75 and 78, and 2 normal days with THI⩽74. Health and behaviour of the bulls were evaluated through 8-h observation sessions starting after morning feed delivery. The study was carried out during a rather cool summer with a climate average THI of 68.9 and 4 days with average THI>75. Despite these mild climate conditions, ceiling fans lowered litter moisture and acted as a preventive measure for bulls' dirtiness (odd ratio=47.9; 95% CI 19.6 to 117.4). The risk of abnormal breathing was increased for Control bulls (odd ratio=40.7; 95% CI 5.4 to 304.2). When exposed to alert THI conditions, respiration rate and panting scores increased and rumination duration dropped in Control bulls compared with bulls provided with a ceiling fan. During observations under alert THI, bulls spent less time eating, more time being inactive and consumed more water compared with normal THI conditions. Bulls' daily dry matter intake measured during the observation sessions decreased on alert compared with normal THI days (P<0.001) due to a drop of intake during the daylight hours. Ceiling fan treatment had no effect on bulls' growth performance or water consumption but these results most likely depended on the mild climate conditions. Ceiling fans proved to mitigate some of the negative effects of heat stress on bulls' behaviour (rumination, lying down and drinking water) and respiration rate, however. The lack of a significant improvement of bulls' growth response should not discourage beef farmers from using ceiling fans in indoor systems, considering the likely increase in frequency and intensity of heat waves in the planet's temperate areas induced by global warming.

von Willebrand disease type 1 mutation p.Arg1379Cys and the variant p.Ala1377Val synergistically determine a 2M phenotype in four Italian patients.

INTRODUCTION: We characterized five patients affected with von Willebrand disease (VWD) carrying the p.Arg1379Cys mutation. One was diagnosed as VWD type 1 and four as type 2M. The 2M patients also have the variant p.Ala1377Val in cis with p.Arg1379Cys. AIM: To evaluate the role of p.Ala1377Val and p.Arg1379Cys von Willebrand factor (VWF) variants to explain patients' phenotype. METHODS: Conventional phenotype tests were used to evaluate patients' plasma and platelets. Direct sequence analysis of exon 28 was carried out. The allele frequency of p.Ala1377Val was evaluated using online database. pcDNA3.1-VWF-WT and mutant (A1377V, R1379C and A1377V-R1379C) expression vectors were transiently transfected in HEK293 cells. The capacity of WT and mutant recombinant (r)VWF (along with patients' plasma VWF) to bind glycoprotein Ibα (GpIbα) were evaluated, using two ELISA assays. One with a wild-type (WT) recombinant (r)GpIbα at increasing ristocetin concentrations (from 0 to 1.50 mg mL-1 ) and the other with a gain-of-function mutant rGpIbα (VWF:GPIbM). RESULTS: The substitution c.4130C>T (p.Ala1377Val) was reported as rare variant in online databases. At 0.25 mg mL-1 of ristocetin, WT, A1377V and R1379C showed 6, 7.5 and 12-fold increased binding to rGpIbα, respectively. A1377V-R1379C rVWF showed no increased binding to rGpIbα at the same ristocetin concentration and reached the highest binding, of only 3-fold increased, at 1.50 mg mL-1 of ristocetin. The VWF:GPIbM showed strongly reduced values for the A1377V-R1379C rVWF and the 2M patients' plasma. CONCLUSION: Our study showed that the presence of both p.Ala1377Val and p.Arg1379Cys mutations (synergistic effect) abolishes the binding of rVWF to rGpIbα, explaining patients' 2M phenotype.

Short communication: Reference limits for blood analytes in Holstein late-pregnant heifers and dry cows: Effects of parity, days relative to calving, and season.

Reference limits for metabolic profiles in Holstein late-pregnant heifers and dry cows were determined considering the effects of parity, days relative to calving, and season. Blood samples were collected from 104 pregnant heifers and 186 dry cows (68 primiparous and 118 pluriparous) from 60 to 10 d before the expected calving date in 31 dairy farms in northeastern Italy. Sampling was performed during summer (182 samples) and the following winter (108 samples). All the animals were judged as clinically healthy at a veterinary visit before sampling. Outliers were removed from data of each blood analyte, and variables that were not normally distributed were log transformed. A mixed model was used to test the fixed effects of parity (late-pregnant heifers, primiparous or pluriparous dry cows), class of days relative to calving (60-41 d, 40-21 d, 20-10 d), season (summer or winter), and the interactions between parity and class of days relative to calving and between parity and season, with farm as random effect. Single general reference limits and 95% confidence intervals were generated for analytes that did not vary according to fixed effects. Whenever a fixed effect included in the model significantly affected a given analyte, specific reference limits and 95% confidence intervals were generated for each of its levels. Albumin, urea, triglycerides, alanine aminotransferase, aspartate aminotransferase, creatinine kinase, conjugated bilirubin, calcium, phosphorus, magnesium, potassium, chloride, zinc, copper, and iron concentrations were not influenced by any of the fixed effects. Total protein, globulins, creatinine, glucose, alkaline phosphatase, gamma glutamyltransferase, lactate dehydrogenase, and sodium plasma concentrations were affected by parity. The class of days relative to calving had a significant effect on the concentrations of total protein, globulins, fatty acids, cholesterol, total bilirubin, and sodium. Season affected plasma concentrations of creatinine, glucose, fatty acids, lactate dehydrogenase, and sodium. Interactions between parity and class of days relative to calving and between parity and season did not significantly affect any of the blood analytes tested. The reference limits and the 95% confidence intervals for blood analytes determined in the study could help dairy practitioners to improve the accuracy of metabolic profile interpretation in Holstein late-pregnant cattle.

Synthetic rubber surface as an alternative to concrete to improve welfare and performance of finishing beef cattle reared on fully slatted flooring.

The aim of this study was to compare a fully slatted concrete floor (concrete slatted (CS)) with the same floor on which synthetic rubber slats were placed on the concrete slats (rubber slatted (RS)) as housing solution for finishing beef cattle. The present study involved five commercial beef cattle farms in which the floor of at least three pens was kept as fully slatted, and in an equal number of pens a rubber cover was placed on the floor, tightly matching the gap profile of the concrete slats to allow the drainage of manure. A total of 326 finishing beef bulls were used (153 on CS and 173 on RS), and regardless of the floor treatment animals were housed in groups of 6 to 12 bulls/pen with a space allowance of 3.1 ± 0.2 m2/bull. Bulls had similar initial live weights (422.3 kg on CS and 425.0 kg on RS), but bulls on RS were heavier at the end of the finishing period with a higher average daily gain than bulls kept on CS (1.53 v. 1.46 kg/day; P<0.05). The proportion of bulls treated for locomotor problems was lower in RS pens compared with CS. Rubber covering prevented the occurrence of bursitis, but it increased the odds for hoof overgrowth at end of the finishing period. Hoof overgrowth detected in vivo in bulls on RS was confirmed at the slaughterhouse by the longer dorsal wall and diagonal lengths of the hoof as well as by a more acute toe angle. Compared with bulls on CS, bulls on RS showed less inactivity and resting time, increased social interactions, decreased abnormal lying down and unsuccessful attempts to lie down, as well as shortened the time for lying down. Bulls in RS pens were dirtier compared with those in CS pens, likely due to the draining gaps being reduced to 11.6 ± 1.2% of the total pen surface compared with the 16.9 ± 1.7% in CS pens. This study gave further evidence about the positive effects of the RS floor on growth performance and welfare of finishing beef cattle, although compromising cleanliness and hoof overgrowth.

Evaluation of an heterogeneous group of patients with von Willebrand disease using an assay alternative to ristocetin induced platelet agglutination.

BACKGROUND: Diagnosis of von Willebrand disease (VWD) type 2 usually relies on the discrepancy between the von Willebrand factor (VWF) ristocetin cofactor activity (VWF:RCo) and VWF antigen (VWF:Ag). Type 2B patients can be discriminated from other qualitative VWD variants by using ristocetin-induced platelet agglutination (RIPA) test. The major limitation of RIPA is the requirement of fresh blood sample. OBJECTIVES: In this study, we evaluated the VWF gain-of-function mutant GPIb binding (VWF:GPIbM) and VWF:RCo assays to investigate whether the VWF:GPIbM/VWF:RCo ratio was able to identify the type 2B variant among an heterogeneous VWD population, previously characterized following the ISTH-SSC guidelines. PATIENTS/METHODS: Seventy-six VWD patients and 31 healthy subjects were evaluated by using VWF:Ag, VWF:RCo, and VWF:GPIbM assays. RESULTS: The mean (minimum-maximum values) VWF:GPIbM/VWF:RCo ratio was higher in type 2B patients (2.53, 0.84-6.11) than in healthy controls (1.05, 0.87-1.34), type 1 (0.85, 0.51-1.15), 2A (1.20, 0.36-2.82), and 2M (1.07, 0.91-1.38) (P < 0.0001). Type 2B variants were divided into four groups (A, B, C, and D) according to their different multimeric patterns. The mean value of the VWF:GPIbM/VWF:RCo ratio in the four groups showed an increasing trend from group A (1.08) to D (3.69), proportional to the loss of high molecular weight multimers. Among 32 type 2B patients, previously diagnosed with RIPA, 8 (mainly with a type I New York/Malmö phenotype) were not confirmed using the VWF:GPIbM/VWF:RCo ratio. CONCLUSIONS: Whenever the RIPA test is not feasible, the VWF:GPIbM/VWF:RCo ratio might help to identify severe type 2B VWD patients.

Robot-assisted radical prostatectomy: recent advances.

Robot assisted radical prostatectomy (RARP) represents the leading application of robotic surgery in the urologic field and it has become the main treatment option for localized prostate cancer (PCa) in the USA In the present review we summarized and critically analyzed the literature of the past five years about this widely used robotic procedure. RARP has continuously evolved in terms of technical modifications and procedural steps. Long-term data are now available, suggesting comparable oncological outcomes to those of open and laparoscopic radical prostatectomy. Good functional outcomes have also been demonstrated. Technological innovations and the introduction of more advanced robotic platforms featuring novel arm-integrated equipment, together with a mature clinical experience with the robotic approach, are likely to lead towards optimal outcomes. Despite the expanding clinical implementation of RARP in the management of prostate cancer, some issues related to this procedure remain matter of debate, such as costs, comparative outcomes versus other approaches, and its role in high risk disease.

Assessment of welfare of finishing beef cattle kept on different types of floor after short- or long-term housing.

This study aimed at evaluating short- and long-term effects of housing beef cattle on deep litter (DL) or concrete fully slatted floor (FS) on their welfare. Animal-based measures of the Welfare Quality® assessment protocol for cattle were used to assess health status and behaviour of bulls. The assessment was carried out in a large commercial farm on 15 batches of bulls (4 DL and 11 FS) 1 month after their receiving day (short-term) and on 12 batches (three DL and nine FS) the week before slaughter (long-term). Signs of better comfort on deep litter in terms of shorter lying down durations (5.1 ± 0.5 v. 6.5 ± 0.4 s; P<0.05) and lower risk of hairless patches (odds ratio=0.09; 95% confidence interval=0.01 to 0.68; P<0.05) were already observed after 1 month. Heavy bulls after a long-term housing on FS showed a higher prevalence of bursitis, hairless patches and lesions/swellings than animals on DL. Bulls on fully slatted floor were at higher risk of early culling (odds ratio=6.44; 95% confidence interval=1.57 to 26.37; P<0.01), mainly due to musculoskeletal system pathologies/lameness. Deep litter proved to be a valid alternative to slatted floor, making animals more confident to interact with powerful movements such as mounting at the end of the finishing period. A negative aspect of the deep litter was the poor cleanliness of the bulls. Compared with the fully slatted floor, there were higher odds ratios for dirty bulls at both, the short- (odds ratio=25.09; 95% confidence interval=8.96 to 70.22; P<0.001) and the long-term housing (odds ratio=276.13; 95% confidence interval=98.21 to 776.38; P<0.001). In order to improve health and welfare of beef cattle finished at a heavy weight, deep litter systems are a promising alternative to fully slatted floors. However, proper management of deep litter is necessary to maintain satisfactory cleanliness of the bulls.