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Miyuki Takashima,1,2 Christian Lalonde,1 Laura Ashley Olszanski,1 Feng-Qi Zhao1 1Department of Animal and Veterinary Sciences, University of Vermont, Burlington, VT, 05405, USA; 2Wakunaga Pharmaceutical Co. Ltd, Osaka, 532-0003, JapanCorrespondence: Feng-Qi Zhao Email fzhao@uvm.eduIntroduction: Milk depression is the major driver of economic loss due to mastitis in dairy animals. The aim of this study was to identify potential mediators of milk depression by investigating the local and systemic changes in gene expression or cytokine production during endotoxin challenge of the mammary gland in a mouse model.Methods: The left and right sides of the 4th pair of mouse mammary glands were alternatively injected with either lipopolysaccharide (LPS, Escherichia coli 055: B5, 50 μL of 0.4 mg/mL) or sterile PBS through the teat meatus 3 days postpartum (n = 9). The 4th glands were individually collected 12 h after LPS injection and analyzed to identify gene expression changes by RNA sequencing and real-time PCR, and the plasma was collected before and after LPS challenge and analyzed to determine the levels of 32 cytokines.Results: Transcriptome analysis showed that in addition to strong pro-inflammatory responses, which included granulocyte and monocyte migration and cytokine production and signaling, the LPS-treated glands exhibited strong ubiquitin-mediated and immune-mediated proteasome activation and an increase in nitric oxide-mediated oxidative stress. Furthermore, LPS induced a down-regulation in vesicle membrane, vesicle-mediated trafficking, and metabolic processes of amino acids and other organic molecules in the mammary gland. Of the 32 cytokines analyzed, the levels of 24 (mainly IL-6, G-CSF, MCP-1, RANTES, MIG, MIP-1b, KC, MIP-2, IP-10, and TNFα) were increased or tended to increase in the blood after LPS treatment, and only the levels of IL-9 were decreased. In the mammary gland after LPS challenge, the levels of IL-5, IL-6, IP-10, LIF, MCP-1, MIP-2, and TNFα were significantly increased, and the levels of INF&Ugr;, IL-2, IL-4, IL-10, and IL-12 (p40) were decreased.Discussion: These observations provide potential markers and targets for further studies on the prevention and treatment of gram-negative bacteria-induced mastitis.Keywords: cytokine, lipopolysaccharide, oxidative stress, proteasome, transcriptome change

Objective: The purpose of this study was to attain consensus among international shoulder experts for the early and appropriate assessment and management strategies of PSI based on classification, recovery timelines, outcome measures and risk. Methods: A three round e-Delphi survey was released over a series of consecutive months to 70 clinical shoulder experts (physiotherapists, orthopaedic surgeons, sport medicine physicians and researchers). Phase 1 consisted of a structured literature review which was integrated with expert input to create 50 items on definitions, classification, assessments, prognosis, and management. The draft Delphi survey was revised through iterative consultation and pilot testing Phase 2 consisted of three rounds of questions; with analysis and survey revision after each round. Consensus was defined as 70% agreement. Descriptive statistics were used to describe the characteristics of the respondents, the response rate of the experts per round and the consensus for PSI classification, assessment, and management. Results: The response rate from each round ranged from 81% in Round 1 to a high of 94% in Round 3. The final survey was completed by 47 individuals from 5 different countries with Canada having the largest representations at 24 (51.1%). Three subgroups of PSI were identified: Traumatic (100% agreement), Microtraumatic (98% agreement) and Atraumatic (98% agreement). After 3 Delphi rounds the agreement of the clinical assessment, risk factors, management and recovery timelines of the PSI sub-groups were established. Conclusion: This Delphi helps to streamline diagnostic subgroups and management strategies for PSI. This may in turn provide framework for future research in the field including randomized-controlled or prospective cohort studies with evidence- informed interventions. Clinical implications: A shoulder expert consensus study on PSI will establish earlier recognition and management strategies for health care providers. [ABSTRACT FROM AUTHOR]