OBJECTIVE Dopamine receptors (DRs) are involved in the development and treatment of many neuropsychiatric disorders. Currently available dopaminergic drugs modulate both DRD2 and DRD3, leading to side effects and uncertainty as to the roles each DR subtype plays physiologically. Our lab employed high throughput screening paradigms to discover highly selective modulators for the DRD3. METHODS The NIH Molecular Libraries Program 400,000 + small molecule library was screened using the Discove RxPathHunter? β- arrestin assay for compounds that activate the DRD3 without effects on the DRD2. Confirmation and counter-screens assessed selectivity and mechanisms of action. We identified 62 potential agonists, and chose the most promising to perform a structure-activity relationship (SAR) study to increase potency while maintaining selectivity. The lead compound identified through this process, ML417, was also characterized using bioluminescence resonance energy transfer (BRET)-based β-arrestin recruitment and G-protein activation assays as well as p-ERK assays. Potential neuroprotective properties of this compound were assessed using a SHSY5Y neuronal cell model. RESULTS ML417 displays potent, DRD3-selective agonist activity in multiple functional assays. Binding and functional GPCR screens (>165 receptors) show ML417 has limited cross-reactivity with other GPCRs. ML417 also displays superior (compared to the reference compound pramipexole),dose-dependent protection against a decrease in neurite length induced by 10 μmol·L-1 of the neurotoxin, 6-hydroxydopamine, in the SHSY5Y cell model. CONCLUSION We have discovered and characterized ML417, a potent and highly selective DRD3 agonist. This compound will be useful as a research tool, and may prove useful as a therapeutic drug lead.

Objective: To investigate anti-multidrug resistance (MDR) activity and safety of the bioactive fraction (CL11) from Codiaeum luzonicum crude leaf extract. Methods: Cytotoxic activity of CL11 against MDR and non- resistant colon cancer cells was assessed using MTT assay. Mode of cell death was investigated by annexin V-propidium iodide staining, TUNEL, and JC-1 assays. To examine mechanism of action, the effect on the expression and function of the MDR-implicated protein P-glycoprotein was tested using Western blotting and calcein assay, respectively. Results: CL11 had an EC 50 of 0.18, 1.03 and 38.52 μg/mL against HCT-15, HCT-15/Dox and HCT116, respectively. Cytotoxicity was mediated by inhibition of P-glycoprotein function and expression. The mode of cell death involved mitochondrial membrane depolarization and was mostly non-apoptotic at EC 50 concentrations against HCT-15 and HCT-15/Dox. Conclusions: Fraction CL11 of Codiaeum luzonicum induces non- apoptotic cell death in MDR cancer cells by overcoming MDR through inhibition of P-glycoprotein expression and function.

Rotational thromboelastometry (ROTEM) is a point-of-care viscoelastic method and enables to assess viscoelastic profiles of whole blood in various clinical settings. ROTEM-guided bleeding management has become an essential part of patient blood management (PBM) which is an important concept in improving patient safety. Here, ROTEM testing and hemostatic interventions should be linked by evidence-based, setting-specific algorithms adapted to the specific patient population of the hospitals and the local availability of hemostatic interventions. Accordingly, ROTEM-guided algorithms implement the concept of personalized or precision medicine in perioperative bleeding management (‘theranostic’ approach). ROTEM-guided PBM has been shown to be effective in reducing bleeding, transfusion requirements, complication rates, and health care costs. Accordingly, several randomized-controlled trials, meta-analyses, and health technology assessments provided evidence that using ROTEM-guided algorithms in bleeding patients resulted in improved patient's safety and outcomes including perioperative morbidity and mortality. However, the implementation of ROTEM in the PBM concept requires adequate technical and interpretation training, education and logistics, as well as interdisciplinary communication and collaboration.
Cooperative Behavior ; Education ; Health Care Costs ; Hemorrhage ; Humans ; Interdisciplinary Communication ; Methods ; Mortality ; Organization and Administration ; Patient Safety ; Point-of-Care Systems ; Precision Medicine ; Technology Assessment, Biomedical ; Thrombelastography

Objective: Agitation in patients diagnosed with personality disorders (PD) is one of the most frequent crises in emergency departments (ED). Although many medications have been tested, their effectiveness has been small or non-significant, and no specific drugs are supported by the available evidence. This study aimed to evaluate the efficacy of Inhaled loxapine (IL) as a therapeutic option for agitated patients with PD. Methods: A naturalistic, unicentric, prospective study was carried out. Thirty subjects diagnosed with PD and attending the ED with episodes of agitation were recruited most of whom were women diagnosed with Borderline Personality Disorder. Subjects were treated with a single dose of IL (9.1 mg). Efficacy was assessed with the Clinical Global Impression scale, the Excited Component of the Positive and Negative Syndrome Scale (PANSS-EC) and the Agitation-Calmness Evaluation Scale (ACES). Patients were followed 60 minutes after administration to measure IL effect and its duration. Results: IL exhibited an overall efficacy in managing mild to severe agitation, with a quick onset of effect and persistence. ‘Effect of time’, where IL efficacy is maintained over time, is more marked in higher-severity agitation. No additional treatments were needed to improve agitation during the follow-up time. Conclusion Results suggest that IL could be a safe and effective option to manage agitation in PD.

Rotational thromboelastometry (ROTEM) is a point-of-care viscoelastic method and enables to assess viscoelastic profiles of whole blood in various clinical settings. ROTEM-guided bleeding management has become an essential part of patient blood management (PBM) which is an important concept in improving patient safety. Here, ROTEM testing and hemostatic interventions should be linked by evidence-based, setting-specific algorithms adapted to the specific patient population of the hospitals and the local availability of hemostatic interventions. Accordingly, ROTEM-guided algorithms implement the concept of personalized or precision medicine in perioperative bleeding management (‘theranostic’ approach). ROTEM-guided PBM has been shown to be effective in reducing bleeding, transfusion requirements, complication rates, and health care costs. Accordingly, several randomized-controlled trials, meta-analyses, and health technology assessments provided evidence that using ROTEM-guided algorithms in bleeding patients resulted in improved patient's safety and outcomes including perioperative morbidity and mortality. However, the implementation of ROTEM in the PBM concept requires adequate technical and interpretation training, education and logistics, as well as interdisciplinary communication and collaboration.