Baseline mutational profiles of patients with carcinoma of unknown primary origin enrolled in the CUPISCO study.

BACKGROUND: Patients with unfavorable carcinoma of unknown primary origin (CUP) have an extremely poor prognosis of ∼1 year or less, stressing the need for more tailored treatments, which are currently being tested in clinical trials. CUPISCO (NCT03498521) was a phase II randomized study of targeted therapy/cancer immunotherapy versus platinum-based chemotherapy in patients with previously untreated, unfavorable CUP, defined as per the European Society for Medical Oncology guidelines. We present a preliminary, descriptive molecular analysis of 464 patients with stringently diagnosed, unfavorable CUP enrolled in the CUPISCO study. MATERIALS AND METHODS: Genomic profiling was carried out on formalin-fixed, paraffin-embedded tissue to detect genomic alterations and assess tumor mutational burden and microsatellite instability. RESULTS: Overall, ∼32% of patients carried a potentially targetable genomic alteration, including PIK3CA, FGFR2, ERBB2, BRAFV600E, EGFR, MET, NTRK1, ROS1, and ALK. Using hierarchical clustering of co-mutational profiles, 10 clusters were identified with specific genomic alteration co-occurrences, with some mirroring defined tumor entities. CONCLUSIONS: Results reveal the molecular heterogeneity of patients with unfavorable CUP and suggest that genomic profiling may be used as part of informed decision-making to identify the potential primary tumor and targeted treatment options. Whether stringently diagnosed patients with unfavorable CUP benefit from targeted therapies in a similar manner to those with matched known primaries will be a key learning from CUPISCO.

Effect of using 200 µg of gonadorelin at the first gonadotropin-releasing hormone of the breeding-Ovsynch on ovulatory response and pregnancies per artificial insemination in first-service lactating Holstein cows.

This study aimed to determine whether 200 µg of GnRH (gonadorelin hydrochloride) would increase ovulatory response and pregnancies per artificial insemination (P/AI) compared with 100 µg at the first GnRH of the breeding-Ovsynch of a Double-Ovsynch program (DO) in lactating Holstein cows. Weekly cohorts of primiparous (n = 719) and multiparous (n = 1,191) cows submitted to DO (GnRH, 7 d later PGF2α, 3 d later GnRH, 7 d later GnRH [G1], 7 d later PGF2α [PG1], 1 d later PGF2α, ∼32 h later GnRH [G2], and ∼16 h later timed artificial insemination [TAI]) for first service, randomly received either 100 µg or 200 µg of GnRH (gonadorelin hydrochloride) at G1 (primiparous, 64-75 DIM; multiparous, 59-70 DIM). Ovulation was determined by ultrasound 2 d after G1 (n = 1,294) and 2 d after G2 (n = 1,020). Blood samples were collected at G1 and at PG1 d to evaluate serum progesterone (P4) concentrations. Conventional (n = 314, Angus; n = 1,084, Holstein) and Holstein sexed semen (n = 276) were used. Pregnancy was diagnosed on d 32, 46, 88, and 200 post-TAI. The high dose of GnRH (200 µg) increased overall ovulatory response to G1 compared with 100 µg (81.3% vs. 65.1%), being similar between parities (primiparous, 72.2%; multiparous, 73.9%). Mean serum P4 concentrations at PG1 did not differ between treatments (100 µg: 9.59 ± 0.15 ng/mL vs. 200 µg: 9.43 ± 0.15 ng/mL). Cows with no ovulation to G1 had higher serum P4 concentrations at G1 than cows with ovulation to G1 (6.27 ± 0.19 ng/mL vs. 4.66 ± 0.07 ng/mL). At PG1, the proportion of cows with functional corpus luteum (98.7% vs. 89.7%) and serum P4 concentrations (9.68 ± 0.12 ng/mL vs. 9.14 ± 0.22 ng/mL) were greater in cows that ovulated to G1 compared with cows that did not ovulate. Also, cows that ovulated to G1 had a greater increase in serum P4 concentrations from G1 to PG1 than cows with no ovulation (5.26 ± 0.12 ng/mL vs. 3.32 ± 0.25 ng/mL). The high dose of GnRH improved overall P/AI at 32 d post-TAI in cows inseminated with conventional semen (54.6% vs. 48.2%) and tended to improve P/AI on 46 (48.8% vs. 44.9%), 88 (47.6% vs. 43.4%), and 200 (45.3% vs. 41.2%) d post-TAI. Primiparous cows inseminated with conventional semen had better P/AI than multiparous cows at d 32 (58.2% vs. 49.4%), 46 (55.1% vs. 44.4%), 88 (53.2% vs. 43.2%) and 200 (51.6% vs. 40.7%) post-TAI. Primiparous cows treated with 200 µg GnRH had lower P/AI on d 32, 46, 88, and 200 post-TAI when inseminated with sexed semen than with conventional semen. In summary, the higher dose of GnRH at G1 improved ovulatory response and P/AI at d 32 post-TAI and tended to improve P/AI at d 46, 88, and 200 post-TAI in cows inseminated with conventional semen. Moreover, the effect of treatment on P/AI in primiparous cows depended on semen type (conventional vs. sexed semen).

Expression of NEDD4L and ENaC in Urinary Extracellular Vesicles in Pre-eclampsia.

OBJECTIVE: To assess changes in expression of renal epithelial sodium channel (ENaC) and NEDD4L, a ubiquitin ligase, in urinary extracellular vesicles (UEV) of pre-eclamptic women compared to normal pregnant controls. METHODS: Urine was collected from pre-eclamptic women (PE, n = 20) or during normal pregnancy (NP, n = 20). UEV were separated by differential ultracentrifugation. NEDD4L, α-ENaC and γ-ENaC were identified by immunoblotting. RESULTS: There was no difference in the expression of NEDD4L (p = 0.17) and α-ENaC (p = 0.10). PE subjects showed increased expression of γ-ENaC by 6.9-fold compared to NP (p < 0.0001). CONCLUSION: ENaC expression is upregulated in UEV of pre-eclamptic subjects but was not associated with changes in NEDD4L.

EPIC: an evaluation of the psychological impact of early-phase clinical trials in cancer patients.

BACKGROUND: Anxiety and depression in patients with cancer is associated with decreased quality of life and increased morbidity and mortality. However, these are often overlooked and untreated. Early-phase clinical trials (EPCTs) recruit patients with advanced cancers who frequently lack future treatment options, which may lead to increased anxiety and depression. Despite this, EPCTs do not routinely consider psychological screening for patients. PATIENTS AND METHODS: This prospective observational study explored levels of anxiety and depression alongside impact of trial participation in the context of EPCTs. The Hospital Anxiety and Depression Scale and the Brief Illness Perceptions Questionnaire were completed at the point of EPCT consent, the end of screening and at pre-specified time points thereafter. RESULTS: Sixty-four patients (median age 56 years; median Eastern Cooperative Oncology Group performance status 1) were recruited. At consent, 57 patients returned questionnaires; 39% reported clinically relevant levels of anxiety whilst 18% reported clinically relevant levels of depression. Sixty-three percent of patients experiencing psychological distress had never previously reported this. Males were more likely to be depressed (P = 0.037) and females were more likely to be anxious (P = 0.011). Changes in anxiety or depression were observed after trial enrolment on an individual level, but not significant on a population level. CONCLUSIONS: Patients on EPCTs are at an increased risk of anxiety and depression but may not seek relevant support. Sites offering EPCTs should consider including psychological screening to encourage a more holistic approach to cancer care and consider the sex of individuals when tailoring psychological support to meet specific needs.

Longitudinal In Vitro Effects of Silver Diamine Fluoride on Early Enamel Caries Lesions.

This laboratory study evaluated the longitudinal surface microhardness changes in early, incipient, noncavitated, white-spot, enamel caries lesions treated with silver diamine fluoride (SDF). Five intervention groups (SDF, AgNO3, KF, 5% sodium fluoride varnish (FV), deionized water (DI)) × two-time intervals after intervention (immediate & delayed pH-cycling) resulted in 10 groups (n=18). Silver nitrate (AgNO3) and potassium fluoride (KF) groups served as controls to assist in evaluating if remineralization effects were due to the silver or fluoride component in SDF. Early, incipient, noncavitated, white-spot, enamel caries lesions were created in bovine enamel, the extent of demineralization was determined using Vickers surface microhardness (VHNlesion). Intervention treatments were applied. Half the specimens from each group underwent immediate 5-day pH-cycling, and half were stored in an incubator with artificial saliva for 2 weeks before undergoing 5-day pH-cycling. After pH-cycling, lesion hardness was evaluated using VHNpost. Specimens were then exposed to a second demineralization challenge, and lesion softening was evaluated (VHNsecdem). Hardness variables were calculated: ΔVHN = VHNpost - VHNlesion; ΔVHNsecdem = VHNsecdem - VHNpost. Data were analyzed using two-way ANOVA (α=0.05). Immediately cycled, SDF had significantly (p<0.0001) greater remineralization than DI, AgNO3, and FV. All delayed cycling groups had significantly greater remineralization than FV (p<0.0001). Significantly greater remineralization was noted in delayed AgNO3 (p≤0.0001), DI (p=0.0003), and FV (p=0.0006) compared to immediately cycled. After the second demineralization challenge, FV had significantly less surface softening than AgNO3 (p=0.0002), DI (p=0.0003), KF (p=0.0225), and SDF (p=0.0388) intervention groups. No significant difference was found between the pH-cycle timings (p=0.2710). Based the present findings, FV may be better suited than SDF to treat early, incipient, noncavitated, white-spot, enamel caries lesions.

Evaluating automated infrared thermography and vulva exposure tracking as components of an estrus detection platform in a commercial dairy herd.

The primary objective of this study was to develop an automated infrared thermography platform (Estrus BenchMark) capable of measuring skin temperature and tail movements as a means of identifying cows in estrus. The secondary objective was to evaluate the accuracy of Estrus BenchMark to detect estrus compared to in-line milk progesterone (P4) analysis (Herd Navigator System) in a commercial dairy herd managed under a robotic milking system. Data were collected on forty-six cows from 45 to 120 d after calving. Cows were flagged in estrus when milk P4 fell below 5 ng/mL. The Estrus BenchMark true positive estrus alerts (Sensitivity; Se%) were compared to Herd Navigator System estrus alerts at different time-windows (±12 h, ±24 h, ±48 h, and ±72 h) relative to the Estrus BenchMark estrus alerts for all the estrus alerts (AE) and confidence-quality estrus (CQE; >80% quality) alerts identified by Herd Navigator System. The Estrus BenchMark captured skin temperature and tail movements resulting in vulva exposure (left tail movements, LTail; right tail movements, RTail; and pooled tail movements, PTail) for each milking event. Skin temperature tended to increase when the milk P4 concentration (Least-Squares Means ±â€¯SE) dropped for AE (estrus day [d 0]; P4; 3.51 ±â€¯0.05 ng/mL, Skin temperature; 33.31 ±â€¯2.38 °C) compared with d -7 (P4; 20.22 ±â€¯0.73 ng/mL; Skin temperature: 32.05 ±â€¯3.77 °C). The increase in skin temperature, however, was significant in cows with CQE > 80% at d 0 (32.75 ±â€¯0.29 °C) compared to d -7 (31.80 ±â€¯0.28 °C). The prevalence of tail movements to expose vulva was greater (P = 0.01) in AE at d 0 (LTail: 62.50%; PTail; 68.75%; and RTail: 56.25%) compared with d -7 (LTail: 18.75%; PTail: 9.37%: and RTail: 9.37%), and d +4 (LTail: 9.37%; PTail: 9.37%; and RTail: 12.5%). Moreover, the higher prevalence of tail movements at d 0 was observed in cows with CQE > 80% (LTail; 65%, PTail; 80%, and RTail; 70%) compared to those with CQE < 80%. The highest Estrus BenchMark Youden index (YJ; 0.45), diagnostic odds ratio (DOR; 9.04), and Efficiency (0.77) were achieved for AE in a ±48 h window and at ±72 h window for CQE (YJ; 0.66, DOR; 25.29, and Efficiency 0.76) relative to Herd Navigator System estrus alerts. The highest Estrus BenchMark resulted in 58% estrus detection rates for AE and 80% for cows with CQE compared to the Herd Navigator System.

Antibiotic use reduces efficacy of tyrosine kinase inhibitors in patients with advanced melanoma and non-small-cell lung cancer.

BACKGROUND: Antibiotic (ABX) use can reduce the efficacy of immune checkpoint inhibitors and chemotherapeutics. The effect for patients treated with targeted therapies, namely, small-molecule tyrosine kinase inhibitors (TKIs), is less known. PATIENTS AND METHODS: Retrospective data were analysed for TKI-treated patients with advanced melanoma and non-small-cell lung cancer (NSCLC) between January 2015 and April 2017 at The Christie NHS Foundation Trust. Data on demographics, disease burden, lactate dehydrogenase (LDH) level, presence of brain metastases, ECOG performance status (PS) and ABX use were collected. Progression-free survival (PFS) and overall survival (OS) were compared between the ABX+ group (ABX within 2 weeks of TKI initiation-6 weeks after) and the ABX- group (no ABX during the same period). RESULTS: A total of 168 patients were included; 89 (53%) with NSCLC and 79 (47%) with melanoma. 55- (33%) patients received ABX. On univariable analysis, ABX+ patients demonstrated shorter PFS (208 versus 357 days; P = 0.008) and OS (294 versus 438 days; P = 0.024). Increased age, poorer PS and higher LDH were associated with shorter PFS and OS. On multivariable analysis, ABX use was independently associated with shorter PFS [hazard ratio (HR) 1.57, 95% confidence interval (CI) 1.05-2.34, P = 0.028] and OS (HR 2.19, 95% CI 1.44-3.32, P = 0.0002). The negative impact of ABX on OS was particularly pronounced for patients with PS of ≥2 (HR 3.82, 95% CI 1.18-12.36, P = 0.025). CONCLUSION: For patients treated with TKIs, ABX use is independently associated with reduced PFS and OS and judicious use is warranted, particularly in patients with poorer PS.

Longevity of Crown Margin Repairs Using Glass Ionomer Cement: A Retrospective Study.

OBJECTIVE: The objective of this study was to determine the survival time of crown margin repairs (CMRs) with glass ionomer and resin-modified glass ionomer cements on permanent teeth using electronic dental record (EDR) data. METHODS: We queried a database of EDR (axiUm; Exan Group, Coquitlam, BC, Canada) in the Indiana University School of Dentistry (IUSD), Indianapolis, IN, USA, for records of patients who underwent CMRs of permanent teeth at the Graduate Operative Dentistry Clinic. Two examiners developed guidelines for reviewing the records and manually reviewed the clinical notes of patient records to confirm for CMRs. Only records that were confirmed with the presence of CMRs were retained in the final dataset for survival analysis. Survival time was calculated by Kaplan-Meier statistics, and a Cox proportional hazards model was performed to assess the influence of age, gender, and tooth type on survival time (a<0.05). RESULTS: A total of 214 teeth (115 patients) with CMR were evaluated. Patient average age was 69.4 ± 11.7 years old. Posterior teeth accounted for 78.5% (n=168) of teeth treated. CMRs using glass ionomer cements had a 5-year survival rate of 62.9% and an annual failure rate (AFR) of 8.9%. Cox proportional-hazards model revealed that none of the factors examined (age, gender, tooth type) affected time to failure. CONCLUSION: The results indicate the potential of CMRs for extending the functional life of crowns with defective margins, thus reducing provider and patient burden of replacing an indirect restoration. We recommend future studies with a larger population who received CMR to extend the generalizability of our findings and to determine the influence of factors such as caries risk and severity of defects on survival time.

Evaluation of infrared thermography combined with behavioral biometrics for estrus detection in naturally cycling dairy cows.

Low estrus detection rates (>50%) are associated to extended calving intervals, low economic profit and reduced longevity in Holstein dairy cows. The objective of this study was to evaluate the accuracy of infrared thermography and behavioral biometrics combined as potential estrus alerts in naturally (not induced) cycling dairy cows housed in a tie-stall barn. Eighteen first lactation cows were subjected to transrectal ultrasonography to determine spontaneous ovulation. The dominant follicle (DF) disappearance was used retrospectively as an indirect indicator of ovulation, and to establish the estrus period (48-24 h prior the DF disappearance). Raw skin temperature (Raw IR) and residual skin temperature (Res IR) were recorded using an infrared camera at the Vulva area with the tail (Vtail), Vulva area without the tail (Vnotail), and Vulva's external lips (Vlips) at AM and PM milking from Day 14 until two days after ovulation was confirmed. Behavioral biometrics were recorded on the same schedule as infrared scan. Behavioral biometrics included large hip movements (L-hip), small hip movements (S-hip), large tail movements and small tail movements to compare behavioral changes between estrus and nonestrus periods. Significant increases in Raw IR skin temperature were observed two days prior to ovulation (Vtail; 35.93 ±â€¯0.27 °C, Vnotail; 35.59 ±â€¯0.27 °C, and Vlips; 35.35 ±â€¯0.27 °C) compared to d -5 (Proestrus; Vtail; 35.29 ±â€¯0.27 °C, Vnotail; 34.93 ±â€¯0.31 °C, and Vlips; 34.68 ±â€¯0.27 °C). No significant changes were found for behavioral parameters with the exception of S-hip movements, which increased at two days before ovulation (d -2; 11.13 ±â€¯1.44 Events/5min) compared to d -5 (7.30 ±â€¯1.02 Events/5min). To evaluate the accuracy of thermal and behavioral biometrics, receiver operating characteristic curve analysis was performed using Youden index (YJ), diagnostic odds ratio, positive likelihood ratio (LR+), Sensitivity, Specificity and Positive predicted value to score the estrus alerts. The greatest accuracy achieved using thermal parameters was for Res IR Vtail PM (YJ = 0.34) and L-hip PM (YJ = 0.27) for behavioral biometrics. Combining thermal and behavioral parameters did not improve the YJ index score but reduced the false-positive occurrence observed by increasing the diagnostic odds ratio (26.62), LR+ (12.47), Specificity (0.97) and positive predicted value (0.90) in a Res IR Vtail PM, S-hip AM, S-hip PM combination. The combination of thermal and behavioral parameters increased the accuracy of estrus detection compared to either thermal or behavioral biometrics, independently in naturally cycling cows during milking.

Detection of Hepatitis A Virus in Strawberries Implicated in an Outbreak in the USA in 1997.

Hepatitis A virus (HAV) was detected in frozen strawberries which had been implicated in a large outbreak of hepatitis A in 1997. The sample was analysed after over 20 years of storage, following a standard method not available at the time of the outbreak. This is the first study in which the HAV associated with the 1997 outbreak of foodborne hepatitis has finally been detected.

Combined effects of soft drinks and nicotine on Streptococcus mutans metabolic activity and biofilm formation.

The purpose of this study was to explore the effects of nicotine on the activity of Streptococcus mutans (S. mutans) in soft drinks. Regular soft drinks contain large proportions of high-fructose corn syrup (HFCS), which increases the activity of S. mutans resulting in high-caries risk compared with sugar-free soft drinks. Nicotine use exhibits a strong correlation with increased S. mutans biofilm formation. The soft drinks chosen were (Coca-Cola Classic, Diet Coke, Coca-Cola Zero Sugar, Caffeine-Free Coca-Cola, Caffeine-Free Diet Coke, Caffeine-Free Coca-Cola Zero Sugar). S. mutans was grown overnight in tryptic soy broth; nicotine was diluted in tryptic soy broth supplemented with 1.0% sucrose followed by soft drinks in dilution of 1:3. Total growth absorbance and biofilm growth were determined by spectrophotometry, absorbance measured to determine biofilm formation, and metabolic activity quantified. One-way ANOVA showed a considerable effect for HFCS and caffeine in the presence of nicotine and their interaction in all measures. Results showed sugar-free caffeinated colas demonstrated significant effect in inhibiting S. mutans biofilm formation and metabolic activity with nicotine. Nicotine-induced S. mutans increased biofilm formation and metabolic activity in the presence of HFCS and caffeine in soft drinks. In conclusion, smokers should consider sugar-free caffeinated versions to minimize the chance of developing dental caries dut to the reduction of biofilm formation.

Clinical evaluation of a universal adhesive in non-carious cervical lesions.

OBJECTIVES: To compare the clinical performance of a universal adhesive in class V non-carious cervical lesions (NCCLs) using two surface treatment protocols (self-etch [SfE] vs selective-enamel-etch [SelE]). MATERIAL AND METHODS: Thirty-three adults, each with ≥2 NCCLs, received one resin composite restoration utilizing a SfE universal adhesive and another utilizing the adhesive and SelE with 37% phosphoric acid. Restorations were evaluated for sensitivity, retention, marginal discoloration, marginal adaptation, and clinical acceptability through 24 months using Cochran-Mantel-Haenszel tests for stratified, ordered categorical outcomes. RESULTS: Sixty-six restorations (35 SfE, 31 SelE; 27 volunteers) were evaluated at 24 months. There were no significant differences between SfE and SelE for sensitivity, retention, marginal adaptation, or clinical acceptability. One SfE restoration was lost. Marginal adaptation was significantly worse at 24 months than baseline for SelE (P = 0.01), but not for SfE. Marginal discoloration was significantly worse for SfE (P = 0.02), but not for SelE. Sensitivity improved from baseline to 24 months for both groups (SelE P = 0.004, SfE P = 0.002). CONCLUSIONS: Twenty-four-month data indicated significantly reduced sensitivity for both groups, worse marginal discoloration for SfE, and worse marginal adaptation for SelE. No changes in retention or clinical acceptability were observed in either group. All retained restorations were clinically acceptable at 24 months. CLINICAL SIGNIFICANCE: Both self-etch and selective enamel etch techniques with a universal adhesive produced clinically acceptable results in resin composite restorations for NCCLs over 2 years.

Chlorhexidine-modified nanotubes and their effects on the polymerization and bonding performance of a dental adhesive.

OBJECTIVES: The purpose of this study was to synthesize chlorhexidine (CHX)-encapsulated aluminosilicate clay nanotubes (Halloysite®, HNTs) and to incorporate them into the primer/adhesive components of an etch-and-rinse adhesive system (SBMP; Scotchbond Multipurpose, 3M ESPE) and to test their effects on degree of conversion, viscosity, immediate and long-term bonding to dentin. METHODS: CHX-modified HNTs were synthesized using 10% or 20% CHX solutions. The primer and the adhesive components of SBMP were incorporated with 15wt.% of the CHX-encapsulated HNTs. Degree of conversion (DC) and viscosity analyses were performed to characterize the modified primers/adhesives. For bond strength testing, acid-etched dentin was treated with one of the following: SBMP (control); 0.2%CHX solution before SBMP; CHX-modified primers+SBMP adhesive; SBMP primer+CHX-modified adhesives; and SBMP primer+CHX-free HNT-modified adhesive. The microtensile bond strength test was performed after immediate (24h) and long-term (6 months) of water storage. Data were analyzed using ANOVA and Tukey (α=5%) and the Weibull analysis. RESULTS: DC was greater for the CHX-free HNT-modified adhesive, whereas the other experimental adhesives showed similar DC as compared with the control. Primers were less viscous than the adhesives, without significant differences within the respective materials. At 24h, all groups showed similar bonding performance and structural reliability; whereas at the 6-month period, groups treated with the 0.2%CHX solution prior bonding or with the CHX-modified primers resulted in greater bond strength than the control and superior reliability. SIGNIFICANCE: The modification of a primer or adhesive with CHX-encapsulated HNTs was an advantageous approach that did not impair the polymerization, viscosity and bonding performance of the materials, showing a promising long-term effect on resin-dentin bonds.

The association of circulating prepartum metabolites, minerals, cytokines and hormones with postpartum health status in dairy cattle.

The objectives were to evaluate differences in circulating prepartum metabolites, minerals, cytokines and hormones based on postpartum disease category and determine critical circulating concentrations of prepartum analytes associated with postpartum disease in 229 cattle from 11 commercial dairies in Alberta, Canada. Blood was collected at 8.8 ± 2.1 d prepartum and analyzed for a wide array of analytes. Cattle were categorized as healthy (n = 76) or as having inflammatory (INF; n = 28), metabolic (MET; n = 34) or inflammatory and metabolic (INFMET; n = 91) postpartum diseases. The prepartum circulating concentrations of Cu were lesser (0.84 vs. 0.90 µg/mL; P = 0.02) and concentrations of Mo (19.1 vs. 16.5 ng/mL; P = 0.04) and NEFA (0.27 vs. 0.18 mmol/L; P = 0.01) were greater in INFMET cattle compared with healthy cattle. The critical threshold for Cu, Mo and NEFA prepartum concentration that predicted INFMET was ≤ 0.81 µg/mL (sensitivity 45.5% and specificity 74.3%), ≥ 9.91 ng/mL (sensitivity 70.0% and specificity 52.7%) and ≥ 0.19 mmol/L (sensitivity 62.2% and specificity 79.7%), respectively. Regardless of differences in the prepartum circulating concentrations of Cu, Mo and NEFA among healthy cattle and those with postpartum disease, the use of these analytes to predict the incidence of postpartum diseases was limited.

Next-generation sequencing as a screening tool for foodborne pathogens in fresh produce.

Next generation sequencing (NGS) approaches are increasingly applied to tracing microbial contaminants entering the food chain due to NGS' untargeted nature and ability to investigate non-culturable (and/or difficult to culture) organisms while yielding genomic information about the microbiota. So far, a plethora of microbes has been shown to be associated with fresh produce, but few studies have utilised NGS to identify contamination with human pathogens. This study aims to establish the limit of detection (LoD) for Salmonella and phage MS2 (a Norovirus surrogate) contamination of fresh produce employing NGS approaches on the Illumina MiSeq: 16S amplicon-sequencing, and RNA-seq, using ScriptSeq (Illumina) and NEBNext (New England BioLabs) kits. ScriptSeq proved the most sensitive approach; delivering an LoD of 104 CFU reaction-1 (Colony Forming Units) for Salmonella and 105 PFU reaction-1 (Plaque Forming Units) for phage MS2. Use of the NEBNext kit resulted in detection of Salmonella at 106 CFU reaction-1 and phage MS2 at 107 PFU reaction-1. 16S amplicon-sequencing yielded a similar LoD of 105 CFU reaction-1 for Salmonella but could not detect MS2. The tested NGS methodologies, in combination with bioinformatics approaches applied, proved less sensitive than conventional microbial detection approaches.

Symposium review: The impact of management and facilities on cow culling rates.

This symposium review examines the association between comfort and cow longevity, with a particular emphasis on optimizing resting behavior in confinement-housed systems. Housed dairy cattle demonstrate a variety of negative behavioral and physiological effects when lying time is restricted, with cows prioritizing the recovery of rest over feeding when both are deprived. There is, however, wide individual-cow variation in daily lying times, influenced by an array of cow-, housing-, and management-related factors. Cow-related factors include individual preference, parity, stage of lactation cycle, milk yield, ill health, and lameness. Lying time tends to increase with age and days in milk and during periods of ill health, whereas milk yield is negatively correlated with lying time. The effect of lameness is complicated by severity and by interactions with bedding type, which modifies the cows' ability to rise and lie down. Generally, lame cows suffer prolonged lying bouts of greater variability in length and take fewer bouts per day. Often this results in an overall increase in lying time. Thus, higher standards of cow comfort and improved cow health are not always reflected by longer lying times. Housing and management factors that influence resting behavior include the design of the resting area, access to the resting space, and the thermal microenvironment of the lying area. Provision of dry, deep loose bedding, stocking cows to allow each animal access to a resting space, allowing sufficient time to access the resting area, and providing heat abatement to reduce heat load optimize resting behavior. Because lameness and poor body condition are commonly found in culled dairy cattle, the link between cow comfort and culling is likely mediated through lameness onset and management. Optimal comfort helps prevent the onset of lameness and facilitates recovery once cows become lame, which limits the effect of lameness on feeding behavior and reduces the risk for other health-related disorders, poor reproductive performance, and early herd removal. Cow comfort cannot be assessed by measuring the duration of lying time alone. Rather, comfort is reflected by the optimization of resting behavior, providing facilities and management to allow cows to lie down when they choose to do so for as long as they need to.

Evaluation of infrared thermography compared to rectal temperature to identify illness in early postpartum dairy cows.

This study evaluated and compared infrared thermography (IRT) and rectal temperature (RT) as screening tests to identify sick transition dairy cows. Holstein cows (n = 72; 42 primiparous) had RT and IRT temperatures taken daily from 1 to 12 days in milk (DIM). Health examinations were performed daily to diagnose retained fetal membrane, milk fever and metritis, and blood was analyzed for ß-hydroxybutyrate at 6 and 9 DIM to diagnose ketosis. Plasma concentrations of cortisol, interleukin-6, tumor necrosis factor α and serum amyloid A at 3, 6, 9 and 12 DIM were included as additional indicators of illness. Cows were categorized as true sick if clinically diagnosed with an illness, or if at least 2 blood parameters were above the normal range. Diagnostic test performances for RT and IRT variables were determined for each variable at a test referent value that provided the highest Youden's (J) index. The best performing screening test depended on the definition of true sickness. In general, the J index for RT was 0.15-0.17 whereas the highest J index for the IRT variables was 0.22 for the mean eye temperature and 0.19 for the mean cheek temperature. Infrared thermography was at least comparable to RT and some IRT variables performed better as a screening tests than RT. Future studies into the automation of IRT for surveillance of early postpartum diseases is warranted.

Advanced schemes for underdense plasma photocathode wakefield accelerators: pathways towards ultrahigh brightness electron beams.

The 'Trojan Horse' underdense plasma photocathode scheme applied to electron beam-driven plasma wakefield acceleration has opened up a path which promises high controllability and tunability and to reach extremely good quality as regards emittance and five-dimensional beam brightness. This combination has the potential to improve the state-of-the-art in accelerator technology significantly. In this paper, we review the basic concepts of the Trojan Horse scheme and present advanced methods for tailoring both the injector laser pulses and the witness electron bunches and combine them with the Trojan Horse scheme. These new approaches will further enhance the beam qualities, such as transverse emittance and longitudinal energy spread, and may allow, for the first time, to produce ultrahigh six-dimensional brightness electron bunches, which is a necessary requirement for driving advanced radiation sources. This article is part of the Theo Murphy meeting issue 'Directions in particle beam-driven plasma wakefield acceleration'.

Quantitative and qualitative assessment of a coding system for reporting CT colonography.

AIM: To validate a coding system implemented to summarise computed tomography colonography (CTC) findings for the detection of suspected colorectal cancer (CRC) by assessing interobserver variability and also to evaluate any weaknesses through qualitative analysis. MATERIALS AND METHODS: All CTC investigations over a 6-month period (01/07/2016 to 31/12/2016) were analysed retrospectively. Each study was read initially by an advanced practitioner radiographer with a final report issued by a consultant gastrointestinal radiologist. Rates of interobserver agreement, using the kappa statistic, provided a quantitative assessment of levels of agreement. Areas of poor interobserver agreement were identified for further qualitative assessment. RESULTS: The present study included 1,321 CTC procedures and the mean age of patients was 68.4 years (range 28-96 years). Percentage agreement for colonic coding was 90% and for extra-colonic coding 47%. This corresponds to kappa scores of 0.69 (substantial agreement) and 0.22 (fair agreement), respectively. Reasons and examples of disagreement in the colonic coding are highlighted. CONCLUSIONS: High interobserver agreement was observed for C coding, suggesting it is a reproducible method of classifying intra-colonic CTC findings. Some of the difference in classifying extra-colonic findings is the perceived importance of incidental findings between readers, as well as differences in skill set; however, some themes recurred in areas of disagreement and recommendations for refining and improving the coding system are provided.

Thermodynamics of standing and lying behavior in lactating dairy cows in freestall and parlor holding pens during conditions of heat stress.

Heat load is a challenge for high-producing dairy cows, with adverse consequences on production, reproduction, and welfare. The objectives of this observational study in 2 commercial dairy herds were to determine the effects of environmental heat stress on standing and lying behavior, to monitor the changes in core body temperature (CBT) of cows during lying and standing bouts, and to compare changes in CBT during standing bouts in freestall pens versus standing in holding pens. High-producing cows were selected for data collection over a period of 6 d of increasing heat stress during a heat wave to which they were not acclimated. A total of 20 cows were fitted with leg accelerometers that recorded lying behavior and with vaginal temperature loggers that recorded CBT. These data were recorded at 30-s intervals. Time in the parlor holding pen was calculated from automated parlor software that recorded milking unit attachment and removal times. Mean daily temperature-humidity index in the pens increased from 68.5 to 79.0 during the 6-d trial, whereas mean daily lying time decreased from 9.5 to 6.2 h/d. The mean number of lying bouts per day remained similar at 11.1 to 12.2, but duration of lying bouts decreased from a high of 49.7 min on the coolest day to 32.8 min on the hottest day. During lying bouts, CBT increased at a mean rate of 0.50°C/h. In contrast, CBT changed at a mean rate of -0.25°C/h when standing in the freestall pens and only -0.09°C/h when standing in the milking-center holding pens. Explanatory models for the CBT at which cows ended either standing or lying bouts were derived from 6 selected lying bouts and 3 selected standing bouts for each cow on each day. The CBT at which a cow ended a lying bout was positively related to CBT and temperature-humidity index at the start of the bout, bout duration, and rate of CBT change during the bout. The CBT at which a cow ended a standing bout was negatively related to bout duration and positively related to start CBT, start temperature-humidity index, and rate of CBT change. Insights into the thermodynamics of standing and lying behavior in dairy cows during periods of heat stress provided by this study may contribute to the development of more effective strategies to mitigate heat load in dairy cattle.