Assessment of sperm morphology is an important part of bull breeding soundness evaluation (BBSE). Whereas the effects ofevaluator experience and evaluation method on the sperm morphology estimation were assessed in other species, no such studywas conducted for bulls. Our objectives were to assess the effects of evaluator experience and number of sperm assessed on BBSEoutcomes. A single eosin-nigrosin sperm morphology slide from individual semen samples, collected from 35 yearling bullswas used. In Experiment I, 6 individuals (3 board-certified theriogenologists [DACT] and 3 fourth-year veterinary students [VS])evaluated 100 sperm from 35 slides twice (at least 1 week between evaluations). In Experiment 2, 3 DACT evaluated 100, 200,and 400 sperm from the same 5 sperm morphology slides to determine if assessing a higher number of sperm would increase theagreement of morphologic characteristics. In Experiment 1, there was a difference (p < 0.0001) in the percent of sperm classifiedas morphologically normal between evaluator types (VS versus DACT). Furthermore, variation among evaluators affected spermmorphology assessments and bull breeding soundness evaluation classifications. Whereas the time needed to evaluate slides increased(p = 0.96) with increasing number of sperm assessed, there was no effect (Experiment 2) of number of sperm evaluatedon percent normal sperm, indicating that evaluating more than 100 sperm may not be justifiable. Further investigation on slidepreparation, microscope use, assessor experience, and continuing education/training is important to ensure the repeatability andvalidity of evaluating bovine sperm morphology.

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Bull sperm morphology assessment varied by evaluator

Introduction: Patient-reported smoking history is frequently used as stratification factor in NSCLC-directed clinical research. However, this classification does not fully reflect the mutational processes in a tumour. Next-generation sequencing can identify mutational signatures associated with tobacco smoking, such as single-base signature 4 (SBS4) and indel-based signature 3 (ID3). This provides an opportunity to redefine the classification of smoking and non-smoking associated NSCLC based on individual genomic tumour characteristics and could contribute to reducing the lung cancer stigma. Methods: Whole genome sequencing data and clinical records were obtained from three prospective cohorts of metastatic NSCLC ( N =316). Relative contributions and absolute counts of SBS4 and ID3 were combined with relative contributions of age-related signatures to divide the cohort into smoking (‘smoking high’) and non - smoking associated (‘smoking low’) clusters. Results: The smoking high ( N =169) and smoking low ( N =147) clusters differed significantly in tumour mutational burden, signature contribution and mutational landscape. This signature-based classification overlapped considerably with smoking history. Yet, 26% of patients with an active smoking history were included in the smoking low cluster, of which 52% harboured an EGFR/ALK/RET/ROS1 alteration, and 4% of patients without smoking history were included in the smoking high cluster. These discordant samples had similar genomic contexts to the rest of their respective cluster. Conclusions : A substantial subset of metastatic NSCLC is differently classified into smoking and non-smoking associated tumours based on smoking-related mutational signatures than based on smoking history. This signature-based classification more accurately classifies patients based on genome-wide context and should therefore be considered as stratification factor in clinical research.

Journal Pre-proof Tobacco smoking related mutational signatures in classifying smoking associated and non-smoking associated non-small cell lung cancer.

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Bull sperm morphology assessment varied by evaluator

Journal Pre-proof Tobacco smoking related mutational signatures in classifying smoking associated and non-smoking associated non-small cell lung cancer.