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Aisha Rafique,1 Anders Etzerodt,2 Jonas H Graversen,3 Søren K Moestrup,3 Frederik Dagnæs-Hansen,2 Holger Jon Møller1 1Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus, Denmark; 2Institute of Biomedicine, Aarhus University, Aarhus, Denmark; 3Department of Molecular Medicine, University of Southern Denmark, Odense, Denmark Background: Vitamin D3 possesses anti-inflammatory and modulatory properties in addition to its role in calcium and phosphate homeostasis. Upon activation, macrophages (M) can initiate and sustain pro-inflammatory cytokine production in inflammatory disorders and play a pathogenic role in certain cancers. Purpose: The main purpose of this study was to encapsulate and specifically target calcitriol to macrophages and investigate the anti-inflammatory properties of calcitriol in vitro and in vivo.Methods: In this study we have designed and developed near-infrared calcitriol PEGylated nanoparticles (PEG-LNP(Cal)) using a microfluidic mixing technique and modified lipid nanoparticles (LNPs) to target the M specific endocytic receptor CD163. We have investigated LNP cellular uptake and anti-inflammatory effect in LPS-induced M in vitro by flow cytometry, confocal microscopy and gene expression analyses. LNP pharmacodynamics, bio-distribution and organ specific LNP accumulation was also investigated in mice in vivo. Results: In vitro, we observed the specific uptake of PEG-LNP(Cal)-hCD163 in human M, which was significantly higher than the non-specific uptake of control PEG-LNP(Cal)-IgG(h) in M. Pre-treatment with encapsulated calcitriol was able to attenuate intracellular TNF- expression, and M surface marker HLA-DR expression more efficiently than free calcitriol in LPS-induced M in vitro. Encapsulated calcitriol diminished mRNA gene levels of TNF-, NF-B, MCP-1 and IL-6, while upregulating IL-10. TNF- and IL-6 protein secretion also decreased. In mice, an in vivo pharmacodynamic study of PEG-LNP(Cal) showed a rapid clearance of IgG and CD163 modified LNPs compared to PEG-LNP(Cal). Antibody modified PEG-LNP(Cal) accumulated in the liver, spleen and kidney, whereas unmodified PEG-LNP(Cal) accumulation was only observed in the liver.Conclusion: Our results show that calcitriol can be effectively targeted to M. Our data confirms the anti-inflammatory properties of calcitriol and this may be a potential way to deliver high dose bioactive calcitriol to M during inflammation in vivo. Keywords: macrophages, 1.25(OH)2D3, calcitriol, lipid nanoparticles, pro-inflammatory cytokines, CD163 targeted drug delivery, gene expression analyses, in vivo pharmacodynamics

Background: Previous studies have reported problems measuring testosterone with immunological assays. Here we explore an automatic second generation immunoassay compared to a LC-MS/MS method., Methods: We collected blood samples from 76 women and measured testosterone, progesterone, gender hormonebinding globulin (SHBG), and albumin employing Cobas e601/c501. Testosterone, androstenedione (andro), dehydroepiandrosterone sulphate (DHEAS), and 17-hydroxyprogesterone (17-OHP) concentrations were measured employing LC-MS/MS. We evaluated the difference between testosterone measured by the two methods and examined the potential interference from the selected steroids and bindings proteins., Results: Testosterone concentrations measured by the two methods yielded: Cobas e601 = 1.240 x (LC-MS/MS) - 0.197, r = 0.84, for testosterone concentrations between 0.22 - 4.9 nmol/L. A positive correlation was observed for the difference between results obtained by the two methods and the sample concentration of DHEAS and andro: Diff (Cobas e601 - LC-MS/MS) = 0.116 x DHEAS - 0.396, r = 0.84 and Diff (Cobas e601 - LC-MS/MS) = 0.08 andro - 0.380, r = 0.58. No statistically significant interference was observed for progesterone, 17-OHP, SHBG, and albumin., Conclusions: We report significant differences between testosterone measurements employing an automatic second generation immunoassay and LC-MS/MS. The difference can be correlated with the measured concentrations of DHEAS and andro, and its magnitude is judged to be of limited clinical relevance. Thus we judge that the automatic second generation immunoassay can be used for routine measurement of testosterone.

Neuroscience based Nomenclature (NbN) is a new system of classifying psychotropic drugs by their pharmacological profile. The NbN was developed to replace the current indication-based nomenclature and to provide an up-to-date and more useful framework to better inform pharmacological decisions. NbN provides updated relevant and specific scientific, regulatory and clinical information, aiming to support rational and lucid prescribing. This pharmacologically driven nomenclature, which highlights pharmacological domains and modes of action, may also increase drug adherence as it clarifies the rationale for selecting a specific psychotropic agent., (Copyright © 2015. Published by Elsevier B.V.)

Current psychopharmacological nomenclature remains wedded in an earlier period of scientific understanding, failing to reflect contemporary developments and knowledge, does not aid clinicians in selecting the best medication for a given patient, and tends to confuse patients by prescribing a drug that does not reflect their identified diagnosis (e.g. prescribe "antipsychotics" to depression). Four major colleges of Neuropsychopharmacology (ECNP, ACNP, Asian CNP, and CINP) proposed a new template comprising a multi-axial pharmacologically-driven nomenclature tested by four surveys. The template has five axes: 1-class (primary pharmacological target and relevant mechanism); 2-family (reflecting the relevant neurotransmitter and mechanism); 3-neurobiological activities; 4-efficacy and major side effects; and 5-approved indications. The results of the surveys suggest that the clinicians found the available indication-based nomenclature system dissatisfactory, non-intuitive, confusing, and doubt-inducing for them and the patients. The proposed five-axis template seeks to upend current usage by placing pharmacology rather than indication as the primary axes, with the proposed nomenclature relating primarily to Axis 1-the class, and usage of the other axes would largely depend upon the extent to which the clinician seeks to deepen the scientific and clinical base of his involvement. A significant proportion of the participants in the four surveys were in favour of the proposed system, a similar number wanted to consider the idea further, and only a small proportion (8.6%) were against it. The proposed five-axis pharmacology based nomenclature template is a system which might refresh and reflect the current scientific concepts of neuropsychopharmacology., (Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.)

Traffic fatalities are a major cause of morbidity and mortality in Iran. Occupational sleep medicine field needs more cost-effective and applicable tests for screening purposes. This study reports on a pilot screening study for drowsy drivers in an urban Iranian sample of commercial drivers. The Maintenance of Wakefulness Test (MWT) measures the ability to remain awake objectively. Sleep latency in MWT is a reasonable predictor of driving simulator performance in drivers. In this study, we evaluate whether the Epworth Sleepiness Scale (ESS) and MWT are equally useful in drivers with possible Excessive Daytime Sleepiness (EDS). 46 consecutive road truck drivers in a transportation terminal entered into this study. The ESS score of patients with normal and abnormal MWT was 3.24±2.4 and 4.08±3 respectively which was not significantly differenced (P value = 0.34). No significant correlation was found between the ESS and sleep latency in MWT (r=-0.28, 95%CI= -0.58 to 0.02). By using the receiver operating characteristic analysis, the area under the curve was found to be 0.57 (95% confidence interval = 0.37- 0.77) which is not statistically acceptable (P value=0.46). Our finding showed that the MWT and ESS do not measure the same parameter.

Background: Epithelial ovarian cancer is one of the leading causes of gynaecological cancer morbidity and mortality in women. Early stage ovarian cancer is usually asymptomatic, therefore, is often first diagnosed when it is widely disseminated. Currently available diagnostics lack the requisite sensitivity and specificity to be implemented as community-based screening tests. The identification of additional biomarkers may improve the diagnostic efficiency of multivariate index assays. The aims of this study were to characterise and compare the ovarian tissue immunohistochemical localisation and plasma concentrations of three putative ovarian cancer biomarkers: human cationic antimicrobial protein-18 (hCAP-18); lactoferrin; and CD163 in normal healthy women and women with ovarian cancer., Methods: In this case-control cohort study, ovarian tissue and blood samples were obtained from 164 women (73 controls, including 28 women with benign pelvic masses; 91 cancer, including 21 women with borderline tumours). Localisation of each antigen within the ovary was assessed by immunohistochemistry and serum concentrations determined by ELISA assays., Results: Immunoreactive (ir) hCAP-18 and lactoferrin were identified in epithelial cells, while CD163 was predominately localised in stromal cells. Tissue ir CD163 increased significantly (P<0.05) with disease grade. Median plasma concentrations of soluble (s)CD163 were significantly greater in the cases (3220 ng/ml) than in controls (2488 ng/ml) (P< 0.01). Median plasma concentrations of hCAP-18 and lactoferrin were not significantly different between cases and controls. The classification efficiency of each biomarker (as determined by the area under the receiver operator characteristic curve; AUC) was: 0.67± 0.04; 0.62 ± 0.08 and 0.51 ± 0.07 for sCD163, hCAP-18 and lactoferrin, respectively. When the 3 biomarkers were modelled using stochastic gradient boosted logistic regression, the AUC increased to 0.95 ± 0.03., Conclusions: The data obtained in this study establishes the localisation and concentrations of CD163, hCAP-18, and lactoferrin in ovarian tumours and peripheral blood. Individually, the 3 biomarkers display only modest diagnostic efficiency as assessed by AUC. When combined in a multivariate index assay, however, diagnostic efficiency increases significantly. As such, the utility of the biomarker panel, as an aid in the diagnosis of cancer in symptomatic women, is worthy of further investigation in a larger phase 2 biomarker trial.

Schizophrenia is a severe mental disorder, characterized by a complex symptomatology and a chronic relapsing course, leading to a more or less intensive residual syndrome. The disorder has strong consequences on social functioning and on the patients' quality of life and is associated with a huge economical burden for the society. The best possible drug treatment should be therefore provided. The introduction of the FGAs was an important step forward, since they were the first specific drug treatment for schizophrenia in the first place. However, the positive therapeutic effect was counterbalanced by problematic side-effects, predominantly different kinds of EPS. The SGAs have a much lower liability to induce EPS, with clozapine inducing no EPS. Additionally, SGAs demonstrated some advantages in terms of a broader spectrum of efficacy, including e.g. negative symptoms. However, for most of the single compounds this efficacy advantage was not as high as initially expected. Thus, there is a need for new developments. Currently, drugs in development are based on the classical transmitter related approaches, following especially the model of clozapine and several other multi-receptor compounds. More and more, the glutamatergic system is of interest in this context as well. Totally new directions, e.g. based on genetic findings or focussing more on brain plasticity are headed for. The question is, whether beside the classical concept of antipsychotics, which covers all symptomatic domains of schizophrenia, compounds demonstrating efficacy only in one syndrome such as e.g. negative symptoms will have a chance in the future development.

Objectives: To investigate the expression of the soluble form of the resident macrophage marker CD163 (sCD163) and its association with core parameters for disease activity, including radiographic progression in early rheumatoid arthritis (RA)., Methods: In a longitudinal sample set from early RA patients (n=34) we measured plasma levels of sCD163 at initiation of treatment and after 9 months of treatment and correlated levels with disease activity in 28 joints (DAS28), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) and total Sharp score (TSS). We also measured plasma levels of sCD163 in 55 healthy volunteers (HV) and in a transverse sample set of chronic (>8 years of disease) RA patients (n=24) and OA patients (n=24) with paired plasma and joint fluid., Results: Early RA patients had significantly higher plasma levels of sCD163 (1.69mg/l (1.42-2.10)) (median (IQR)) at baseline than after 9 months of treatment (1.28mg/l (0.963-1.66), p=0.001), but not significantly changed compared with HV (1.66mg/l (1.22-2.02)). In early RA patients, baseline levels of sCD163, correlated with DAS28, CRP and ESR. Interestingly, sCD163 at 9 months was associated with radiographic progression (TSS) between year 0 and 5 (r=0.468, p=0.02). Levels of sCD163 were higher in RA patients, than in OA patients and higher in SF than in plasma., Conclusions: Plasma levels of macrophage derived sCD163 are associated with disease activity and predict radiographic progression in early RA patients, supporting that sCD163 may have a role as a biomarker of disease activity and that resident macrophages are important for joint destruction.

Sickle cell disease (SCD) is characterized by vasculopathy, which has been causally linked to intravascular haemolysis and high levels of free plasma haemoglobin. Soluble CD163 (sCD163) is implicated in the clearance of free plasma haemoglobin and high plasma concentrations have been linked to arterial disease. We therefore investigated the value of sCD163 as a biomarker in children with SCD, and also measured haptoglobin levels in this population. We measured sCD163 in 25 control children with no haemoglobinopathy, 41 with sickle cell anaemia (HbSS) in the steady state, 27 with HbSS taking hydroxycarbamide, and 7 with HbSC disease. There was no significant difference between sCD163 levels in steady-state HbSS (1·78 mg/l) and controls (1·81 mg/l) (P = 0·86). However, sCD163 levels were significantly lower in those HbSS children taking hydroxycarbamide (1·35 mg/l) compared to both steady state HbSS (P = 0·004) and controls (P = 0·036). In children on hydroxycarbamide, sCD163 correlated negatively and highly significantly with percentage HbF (R = -0·76, P < 0·001), and this relationship was absent in those not taking hydroxycarbamide (R = 0·07, P = 0·65). sCD163 is a potentially useful biomarker in children with SCD, and may have a role in monitoring responses to hydroxycarbamide., (© 2011 Blackwell Publishing Ltd.)

Oscillations in the gamma-band frequency range have been described to be more closely connected to hemodynamic changes as assessed with functional magnetic resonance imaging (fMRI) than other aspects of neuronal activity. In addition, gamma-band oscillations have attracted much interest during the last few years since they are thought to play a crucial role in many aspects of brain function related to perception and cognition. It was the aim of the present simultaneous EEG-fMRI study to identify brain regions specifically involved in the generation of the auditory gamma-band response (GBR) using single-trial coupling of EEG and fMRI. Ten healthy subjects participated in this study. Three different runs of an auditory choice reaction task with increasing difficulty were performed. Brain activity was recorded simultaneously with high density EEG (61 channels) and fMRI (1.5 T). BOLD correlates of the GBR have been predicted using the single-trial amplitude of the GBR. Reaction times (p<0.001), error rates (p<0.05) and self-ratings of task difficulty and effort demands (p<0.001) were related to the level of difficulty in the task. In addition, we found a significant influence of task difficulty on the amplitude of the GBR at Cz (p<0.05). Using single-trial coupling of EEG and fMRI GBR-specific activations were found only in the auditory cortex, the thalamus and the anterior cingulate cortex (ACC) in the most difficult run. Single-trial coupling might be a useful method in order to increase our knowledge about the functional neuroanatomy of "neural ensembles" coupled by 40 Hz oscillations., (Copyright (c) 2009 Elsevier Inc. All rights reserved.)