A nationwide survey on producer and veterinarian perceptions of the painfulness of procedures and disease states in dairy and beef cattle.

Failure to adequately manage pain in cattle causes suffering and is thus a welfare concern for the livestock industry. The objectives of this study were to summarize caregiver perceptions of the painfulness of various procedures and disease conditions in cattle. This survey also assessed factors that impact the perception of painfulness and determined relationships between pain perception and mitigation in producers and veterinarians in the United States beef and dairy cattle industries.

Frequency of disagreements between producers and veterinarians about pain management in cattle.

Pain management is a key element of ensuring animal welfare. Although the opinions of both producers and veterinarians affect decisions about the use of pain mitigation on cattle operations, little is known about how they communicate about this topic. Given the importance of a veterinary-client-patient relationship for developing pain mitigation protocols, understanding the communication between veterinarians and producers is key to the implementation of robust, industry-wide pain management protocols.

GPCR heteromers: An overview of their classification, function and physiological relevance.

G protein-coupled receptors (GPCRs) are capable of interacting to form higher order structures such as homomers and heteromers. Heteromerisation in particular has implications for receptor function, with research showing receptors can attain unique expression, ligand binding, signalling and intracellular trafficking upon heteromerisation. As such, GPCR heteromers represent novel drug targets with extensive therapeutic potential.

Novel Pharmacology Following Heteromerization of the Angiotensin II Type 2 Receptor and the Bradykinin Type 2 Receptor.

The angiotensin type 2 (AT) receptor and the bradykinin type 2 (B) receptor are G protein-coupled receptors (GPCRs) that have major roles in the cardiovascular system. The two receptors are known to functionally interact at various levels, and there is some evidence that the observed crosstalk may occur as a result of heteromerization. We investigated evidence for heteromerization of the AT receptor and the B receptor in HEK293FT cells using various bioluminescence resonance energy transfer (BRET)-proximity based assays, including the Receptor Heteromer Investigation Technology (Receptor-HIT) and the NanoBRET ligand-binding assay.

Orexin Signaling: A Complex, Multifaceted Process.

The orexin system comprises two G protein-coupled receptors, OX and OX receptors (OXR and OXR, respectively), along with two endogenous agonists cleaved from a common precursor (prepro-orexin), orexin-A (OX-A) and orexin-B (OX-B). For the receptors, a complex array of signaling behaviors has been reported. In particular, it becomes obvious that orexin receptor coupling is very diverse and can be tissue-, cell- and context-dependent.

Profiling novel pharmacology of receptor complexes using Receptor-HIT.

Many receptors are able to undergo heteromerisation, leading to the formation of receptor complexes that may have pharmacological profiles distinct from those of the individual receptors. As a consequence of this, receptor heteromers can be classed as new drug targets, with the potential for achieving greater specificity and selectivity over targeting their constituent receptors. We have developed the Receptor-Heteromer Investigation Technology (Receptor-HIT), which enables the detection of receptor heteromers using a proximity-based reporter system such as bioluminescence resonance energy transfer (BRET).

The oncogene AAMDC links PI3K-AKT-mTOR signaling with metabolic reprograming in estrogen receptor-positive breast cancer.

Adipogenesis associated Mth938 domain containing (AAMDC) represents an uncharacterized oncogene amplified in aggressive estrogen receptor-positive breast cancers. We uncover that AAMDC regulates the expression of several metabolic enzymes involved in the one-carbon folate and methionine cycles, and lipid metabolism. We show that AAMDC controls PI3K-AKT-mTOR signaling, regulating the translation of ATF4 and MYC and modulating the transcriptional activity of AAMDC-dependent promoters.

Complex interactions between the angiotensin II type 1 receptor, the epidermal growth factor receptor and TRIO-dependent signaling partners.

Transactivation of the epidermal growth factor receptor (EGFR) by the angiotensin II (AngII) type 1 (AT) receptor is involved in AT receptor-dependent growth effects and cardiovascular pathologies, however the mechanisms underpinning this transactivation are yet to be fully elucidated. Recently, a potential intermediate of this process was identified following the discovery that a kinase called TRIO was involved in AngII/AT receptor-mediated transactivation of EGFR. To investigate the mechanisms by which TRIO acts as an intermediate in AngII/AT receptor-mediated EGFR transactivation we used bioluminescence resonance energy transfer (BRET) assays to investigate proximity between the AT receptor, EGFR, TRIO and other proteins of interest.

Pharmacists' views and desires regarding pharmacist administration of vaccines in New Zealand.

Objectives: To explore pharmacists' views and experiences of pharmacist-administered vaccinations, motivators and barriers to pharmacists administering vaccinations and their preferences for expansions to such services.

Methods: All practising pharmacist members (n = 3400) of the Pharmaceutical Society of New Zealand were invited to participate in an online survey in 2017.

Key Findings: A total of 468 pharmacists completed the survey (14%).

Investigation of Receptor Heteromers Using NanoBRET Ligand Binding.

Receptor heteromerization is the formation of a complex involving at least two different receptors with pharmacology that is distinct from that exhibited by its constituent receptor units. Detection of these complexes and monitoring their pharmacology is crucial for understanding how receptors function. The Receptor-Heteromer Investigation Technology (Receptor-HIT) utilizes ligand-dependent modulation of interactions between receptors and specific biomolecules for the detection and profiling of heteromer complexes.

Producer and Veterinarian Perspectives towards Pain Management Practices in the US Cattle Industry.

Producers and veterinarians are considered responsible for improving animal welfare, as they are responsible for implementing practices that directly impact the animal's well-being. Most husbandry procedures performed in cattle do not include pain mitigation, and understanding challenges faced by these stakeholders to use analgesics is key in improving on-farm pain management strategies. Therefore, the objectives of this study were to explore producer and veterinarian perspectives on pain management practices by (1) exploring inquires received by Food Animal Residue Avoidance Databank (FARAD) regarding analgesic use in cattle and (2) using a survey instrument to identify factors that impact pain management implementation in the US cattle industry.

Current attitudes of veterinarians and producers regarding the use of local and systemic analgesia in beef and dairy cattle in the United States.

Objective: To survey cattle producers and veterinarians about the use of analgesia on US cattle operations.

Sample: 1,187 members of the following database, electronic mailing lists, and social media groups: FarmProgress master file, American Association of Bovine Practitioners, Academy of Veterinary Consultants, National Milk Producers Federation Farm Evaluators, Dairy Moms Facebook group, and Dairy Girl Network Facebook group.

Procedures: An online survey was developed to gather information about the frequency of local and systemic analgesia use for common painful procedures and diseases in cattle < 2, 2 to 12, and > 12 months old.

Insights into the Interaction of LVV-Hemorphin-7 with Angiotensin II Type 1 Receptor.

Hemorphins are known for their role in the control of blood pressure. Recently, we revealed the positive modulation of the angiotensin II (AngII) type 1 receptor (AT1R) by LVV-hemorphin-7 (LVV-H7) in human embryonic kidney (HEK293) cells. Here, we examined the molecular binding behavior of LVV-H7 on AT1R and its effect on AngII binding using a nanoluciferase-based bioluminescence resonance energy transfer (NanoBRET) assay in HEK293FT cells, as well as molecular docking and molecular dynamics (MD) studies.

Honeybee venom and melittin suppress growth factor receptor activation in HER2-enriched and triple-negative breast cancer.

Despite decades of study, the molecular mechanisms and selectivity of the biomolecular components of honeybee () venom as anticancer agents remain largely unknown. Here, we demonstrate that honeybee venom and its major component melittin potently induce cell death, particularly in the aggressive triple-negative and HER2-enriched breast cancer subtypes. Honeybee venom and melittin suppress the activation of EGFR and HER2 by interfering with the phosphorylation of these receptors in the plasma membrane of breast carcinoma cells.

Global Forum on Quality Assurance in CE/CPD: Assuring Quality across Boundaries.

As a result of the globalization of access and provision of continuing education and continuing professional development (CE/CPD), the national CE/CPD accreditation organizations of Australia, Canada, Ireland, New Zealand, South Africa, United Kingdom and United States formed the Global Forum on Quality Assurance of Continuing Education and Continuing Professional Development (GFQACE) to investigate and develop means of recognizing CE/CPD across boundaries. Two priorities were identified at their first meeting in 2016: (1) the development of an accreditation framework and (2) the identification of models and approaches to mutual recognition. The GFQACE approved an accreditation framework and facilitated review approach to mutual recognition in 2018 and is currently working on implementation guides.

AT1R-AT2R-RXFP1 Functional Crosstalk in Myofibroblasts: Impact on the Therapeutic Targeting of Renal and Cardiac Fibrosis.

Background: Recombinant human relaxin-2 (serelaxin), which has organ-protective actions mediated its cognate G protein-coupled receptor relaxin family peptide receptor 1 (RXFP1), has emerged as a potential agent to treat fibrosis. Studies have shown that serelaxin requires the angiotensin II (AngII) type 2 receptor (ATR) to ameliorate renal fibrogenesis and . Whether its antifibrotic actions are affected by modulation of the AngII type 1 receptor (ATR), which is expressed on myofibroblasts along with RXFP1 and ATR, is unknown.

Knowledge and Opinions of Third Year Veterinary Students Relevant to Animal Welfare Before and After Implementation of a Core Welfare Course.

Although leading veterinary organizations emphasize the importance of animal welfare knowledge, there exists a gap in current veterinary student animal welfare education and training. A survey instrument was created to assess third-year Doctor of Veterinary Medicine (DVM) student knowledge of key animal welfare topics, opinions regarding the inclusion of welfare education in the veterinary curriculum, and views on veterinarian responsibilities as advocates. In Spring 2018, Colorado State University added a required animal welfare course to the DVM curriculum.

NanoBRET: The Bright Future of Proximity-Based Assays.

Bioluminescence resonance energy transfer (BRET) is a biophysical technique used to monitor proximity within live cells. BRET exploits the naturally occurring phenomenon of dipole-dipole energy transfer from a donor enzyme (luciferase) to an acceptor fluorophore following enzyme-mediated oxidation of a substrate. This results in production of a quantifiable signal that denotes proximity between proteins and/or molecules tagged with complementary luciferase and fluorophore partners.

Transactivation of RAGE mediates angiotensin-induced inflammation and atherogenesis.

Activation of the type 1 angiotensin II receptor (AT1) triggers proinflammatory signaling through pathways independent of classical Gq signaling that regulate vascular homeostasis. Here, we report that the AT1 receptor preformed a heteromeric complex with the receptor for advanced glycation endproducts (RAGE). Activation of the AT1 receptor by angiotensin II (Ang II) triggered transactivation of the cytosolic tail of RAGE and NF-κB-driven proinflammatory gene expression independently of the liberation of RAGE ligands or the ligand-binding ectodomain of RAGE.

NanoBRET ligand binding at a GPCR under endogenous promotion facilitated by CRISPR/Cas9 genome editing.

Bioluminescence resonance energy transfer (BRET) is a versatile tool used to investigate membrane receptor signalling and function. We have recently developed a homogenous NanoBRET ligand binding assay to monitor interactions between G protein-coupled receptors and fluorescent ligands. However, this assay requires the exogenous expression of a receptor fused to the nanoluciferase (Nluc) and is thus not applicable to natively-expressed receptors.

BRET-based assay to monitor EGFR transactivation by the ATR reveals G protein-independent activation and ATR-EGFR complexes.

The type 1 angiotensin II (AngII) receptor (ATR) transactivates the epidermal growth factor receptor (EGFR), which leads to pathological remodeling of heart, blood vessels and kidney. End-point assays are used as surrogates of EGFR activation, however these downstream readouts are not applicable to live cells, in real-time. Herein, we report the use of a bioluminescence resonance energy transfer (BRET)-based assay to assess recruitment of the EGFR adaptor protein, growth factor receptor-bound protein 2 (Grb2), to the EGFR.

Using nanoBRET and CRISPR/Cas9 to monitor proximity to a genome-edited protein in real-time.

Bioluminescence resonance energy transfer (BRET) has been a vital tool for understanding G protein-coupled receptor (GPCR) function. It has been used to investigate GPCR-protein and/or -ligand interactions as well as GPCR oligomerisation. However the utility of BRET is limited by the requirement that the fusion proteins, and in particular the donor, need to be exogenously expressed.

Bioluminescence Resonance Energy Transfer Approaches to Discover Bias in GPCR Signaling.

Bioluminescence resonance energy transfer (BRET) is a well-established technique for investigating G protein-coupled receptor (GPCR) pharmacology. BRET enables the monitoring of molecular proximity through the use of heterologously expressed proteins of interest and/or fluorophore-labeled ligands. Fusion to a donor luciferase enzyme or an acceptor fluorophore and subsequent detection of resonance energy transfer indicate the close proximity of the molecules of interest.

Application of BRET to monitor ligand binding to GPCRs.

Bioluminescence resonance energy transfer (BRET) is a well-established method for investigating protein-protein interactions. Here we present a BRET approach to monitor ligand binding to G protein-coupled receptors (GPCRs) on the surface of living cells made possible by the use of fluorescent ligands in combination with a bioluminescent protein (NanoLuc) that can be readily expressed on the N terminus of GPCRs.