Recent investigations have indicated that 3D skin models are well suitable for drug testing and preclinical studies of topical therapies. The analysis of skin diseases has recognized the importance of inflammatory mechanisms and immune responses and thus other cell types such as dendritic cells and T cells in the skin. Current developments include the production of more complete skin models comprising a range of different cell types. Organ models and even multi-organ systems are being developed for the analysis of higher levels of cellular interaction and drug responses and are among the most recent innovations in skin modelling. They promise improved robustness and flexibility and aim at a body-on-a-chip solution for comprehensive pharmaceutical in vitro studies.Drug delivery to the brain is challenging to study due to the complexity of the barriers of the central nervous system (CNS). The present chapter describes and compares experimental methods such as microdialysis, two-photon laser scanning fluorescence microscopy and positron emission tomography (PET) that can be used for in vivo studies of drug transport across the blood-brain barrier (BBB). The selection of appropriate method is based on the research question, and the different methods will in most cases provide complementary information. Attention is also given to the fact that the BBB might undergo changes in integrity, protein expression and other morphological alterations as a result of disease. The use of animal models of human disease is therefore also discussed. Special emphasis is given to translational aspects of the different methods and readouts.The kappa opioid receptor (KOR) has emerged as a promising therapeutic target for pain and itch treatment. There is growing interest in biased agonists that preferentially activate select signaling pathways downstream of KOR activation on the cellular level due to their therapeutic promise in retaining the analgesic and antipruritic effects and eliminating the sedative and dysphoric effects of KOR signaling on the physiological level. find more The concept of ligand-selective signaling includes that biased ligands promote KOR to selectively recruit one transducer or regulator protein over another, introducing bias into the signaling cascade at the very receptor-proximal level. Measuring agonist effects directly at the receptor has remained challenging and previous studies have focused on inferring agonist-selective KOR engagement with G protein relative to β-arrestin based on downstream signaling readouts. Here we discuss novel strategies to directly assess ligand-selective effects on receptor activation using KOR-interacting biosensors. The conformation-specific cytoplasmic biosensors are disconnected from the endogenous signaling machinery and provide a direct receptor-proxy readout of ligand effects in living cells. Receptor-biosensor interaction is ligand concentration dependent and can be used to determine relative ligand potency and efficacy. In addition, the biosensors reveal the existence of two dimensions of agonist bias in the cellular context Firstly, agonists can selectively produce discrete protein-engaged KOR states and secondly, agonists can differ in the precise subcellular location at which they activate KOR. We discuss the value and the limitations of using orthogonal receptor-interacting biosensors in the quest to understand functional selectivity amongst KOR agonists in the cellular context.The influenza virus causes severe respiratory illnesses and deaths worldwide every year. It spreads quickly in an overcrowded area like the annual Hajj pilgrimage in Saudi Arabia. Vaccination is the primary strategy for protection against influenza. Due to the occurrence of antigenic shift and drift of the influenza virus, a mismatch between vaccine strains and circulating strains of influenza may occur. The objective of this study is to assess the impact of mismatch between vaccine strains and circulating strains during Hajj, which brings together individuals from all over the globe. To this end, we develop deterministic mathematical models of influenza with different populations and strains from the northern and southern hemispheres. Our results show that the existence and duration of an influenza outbreak during Hajj depend on vaccine efficacy. In this concern, we discuss four scenarios vaccine strains for both groups match/mismatch circulating strains, and vaccine strains match their target strains and mismatch the other strains. Further, there is a scenario where a novel pandemic strain arises. Our results show that as long as the influenza vaccines match their target strains, there will be no outbreak of strain H1N1 and only a small outbreak of strain H3N2. Mismatching for non-target strains causes about 10,000 new H3N2 cases, and mismatching for both strains causes about 2,000 more new H1N1 cases and 6,000 additional H3N2 cases during Hajj. Complete mismatch in a pandemic scenario may infect over 342,000 additional pilgrims (13.75%) and cause more cases in their home countries. Denosumab is a humanized IgG2 monoclonal antibody that was approved for the treatment of osteoporosis in Japan in 2013. This study aimed to investigate the long-term safety and effectiveness of denosumab in Japanese patients with osteoporosis in daily clinical practice. This 3-year, prospective, observational, post-marketing study included patients who initiated treatment with denosumab (60mg/6months) for osteoporosis. Data were assessed at baseline, 3, 6, 12, 24 and 36months. Key endpoints were adverse events (AEs), adverse drug reactions (ADRs), occurrence of osteoporotic fractures, bone mineral density (BMD), and bone turnover markers. Multivariate analyses were conducted to identify predictors of hypocalcaemia and percent change in BMD. Overall, 3534 patients were assessed (mean 75.7years; 89.8% women). In total, 298 patients (8.4%) developed ADRs; the most common was hypocalcaemia (3.9%). Hypocalcaemia risk was significantly increased in patients with creatinine clearance < 30mL/min, no prior use of bisphosphonates, prior use of calcium and vitamin D preparations, baseline serum calcium < 8.5mg/dL, and no concomitant use of calcium or vitamin D preparations. Six patients had adjudicated osteonecrosis of the jaw. Lumbar spine BMD increased significantly from baseline (mean percent change 11.4% at 36months). All bone turnover markers decreased significantly from baseline. Over 3years, 3.3% of patients developed a new osteoporotic fracture. This study confirmed the long-term safety and effectiveness of denosumab in Japanese patients with osteoporosis in daily clinical practice. No new safety signals were identified.This study confirmed the long-term safety and effectiveness of denosumab in Japanese patients with osteoporosis in daily clinical practice. No new safety signals were identified.