Portable video media for presenting informed consent and wound care instructions for skin biopsies: a randomized controlled trial

Background Delayed hypersensitivity reactions to drugs can be life‐threatening and constitute a growing problem in clinical practice. Although drug‐specific T cells seem to be involved, the cellular and molecular bases of their aetiopathology are not fully understood. Objectives To study the molecular mechanisms underlying the pathogenesis and the clinical heterogeneity of cutaneous delayed hypersensitivity reactions to drugs. Materials and methods We characterized the gene expression profiles of peripheral blood mononuclear cells (PBMCs) isolated from patients during the acute phase of the reaction and upon resolution of clinical symptoms using a cDNA array technology. Low‐density arrays were used to confirm differential expression of selected genes during the acute disease in patients and to compare gene expression in patients and exposed control donors by quantitative real‐time polymerase chain reaction. Results Eighty‐five genes were found to be differentially expressed during the acute phase of cutaneous drug‐induced delayed hypersensitivity reactions. Furthermore, 92 genes with distinct expression patterns in severe and benign diseases during the acute phase were identified. PBMCs from patients with severe bullous diseases showed a characteristic gene expression pattern with lower expression of genes encoding T cell‐specific proteins and high expression of cell cycle‐related genes and genes coding for inflammatory‐related mediators among which several endogenous damage‐associated molecular patterns (DAMPs) or alarmins were found. Conclusions Distinct gene expression profiles in PMBCs define benign and severe clinical entities. Overexpression of endogenous DAMPs in Stevens–Johnson syndrome and toxic epidermal necrolysis suggest that drugs can trigger the alarmin system in sensitized patients leading to life‐threatening diseases.     

Characterisation of endogenous players in fibroblast growth factor‐regulated functions of hypothalamic tanycytes and energy‐balance nuclei

The mammalian hypothalamus regulates key homeostatic and neuroendocrine functions ranging from circadian rhythm and energy balance to growth and reproductive cycles via the hypothalamic‐pituitary and hypothalamic‐thyroid axes. In addition to its neurones, tanycytes are taking centre stage in the short‐ and long‐term augmentation and integration of diverse hypothalamic functions, although the genetic regulators and mediators of their involvement are poorly understood. Exogenous interventions have implicated fibroblast growth factor (FGF) signalling, although the focal point of the action of FGF and any role for putative endogenous players also remains elusive. We carried out a comprehensive high‐resolution screen of FGF signalling pathway mediators and modifiers using a combination of in situ hybridisation, immunolabelling and transgenic reporter mice, aiming to map their spatial distribution in the adult hypothalamus. Our findings suggest that β‐tanycytes are the likely focal point of exogenous and endogenous action of FGF in the third ventricular wall, utilising FGF receptor (FGFR)1 and FGFR2 IIIc isoforms, but not FGFR3. Key IIIc‐activating endogenous ligands include FGF1, 2, 9 and 18, which are expressed by a subset of ependymal and parenchymal cells. In the parenchymal compartment, FGFR1‐3 show divergent patterns, with FGFR1 being predominant in neuronal nuclei and expression of FGFR3 being associated with glial cell function. Intracrine FGFs are also present, suggestive of multiple modes of FGF function. Our findings provide a testable framework for understanding the complex role of FGFs with respect to regulating the metabolic endocrine and neurogenic functions of hypothalamus in vivo.     

Hemodynamic Effects of Nitroglycerin Ointment in Emergency Department Patients

Background

Nitroglycerin ointment is commonly used in the treatment of emergency department (ED) patients with suspected acute heart failure (AHF) or suspected acute coronary syndrome (ACS), but its hemodynamic effects in this population are not well described.

Objective

Our objective was to assess the effect of nitroglycerin ointment on mean arterial pressure (MAP) and systemic vascular resistance (SVR) in ED patients receiving nitroglycerin. We hypothesized that nitroglycerin ointment would result in a reduction of MAP and SVR in the acute treatment of patients.

Methods

We conducted a prospective, observational pilot study in a convenience sample of adult patients from a single ED who were treated with nitroglycerin ointment. Impedance cardiography was used to measure MAP, SVR, cardiac output (CO), stroke volume (SV), and thoracic fluid content (TFC) at baseline and at 30, 60, and 120 min after application of nitroglycerin ointment. Mixed effects regression models with random slope and random intercept were used to analyze changes in hemodynamic parameters from baseline to 30, 60, and 120 min after adjusting for age, sex, and final ED diagnosis of AHF.

Results

Sixty-four subjects with mean age of 55 years (interquartile range, 48–67 years) were enrolled; 59% were male. In the adjusted analysis, MAP and TFC decreased after application of nitroglycerin ointment (p = 0.001 and p = 0.043, respectively). Cardiac index, CO, SVR, and SV showed no change (p = 0.113, p = 0.085, p = 0.570, and p = 0.076, respectively) over time.

Conclusions

Among ED patients who are treated with nitroglycerin ointment, MAP and TFC decrease over time. However, other hemodynamic parameters do not change after application of nitroglycerin ointment in these patients.     

Metabolomics in pancreatic cancer biomarkers research

Pancreatic cancer is one of the worst prognoses of all malignancies. More than 40,000 deaths a year from this disease are observed in European Union alone. The only possibly curative treatment of pancreatic cancer is surgery, yet only 15–20% of patients have operable disease and even patients, which go through surgery and adjuvant chemotherapy, survival is less than 30%. The sensitive and specific biomarkers which could be used for the advance of early diagnostics are needed and constantly researched. Metabolomics is a technology which analyzes the concentrations of low-molecular-weight metabolites (the metabolome) has lately effectively developed due to the improvements in analytical technology. Metabolome analysis can be a one of the useful approaches for the biomarker discovery and disease diagnosis. Here we discuss recent discoveries in the field of pancreatic cancer metabolomics as well as the most promising biomarkers for diagnostics, prognosis and prediction.     

Experimental survey of non-clonogenic viability assays for adherent cells in vitro.

Results of rapid cell viability assays were experimentally compared in order to reveal the most suitable test for in vitro investigations of the combination of photodynamic therapy (PDT) with chemotherapeutic drugs. meso-Tetra(3-hydroxyphenyl)-chlorin (m-THPC) accumulating in cell membranes and meso-tetra(4-sulfonatophenyl)-porphin (TPPS4) accumulating in lysosomes were used as photosensitisers. Doxorubicin that localises, mainly, to nucleus and vincristine that binds to microtubules were used as cytostatic drugs. Two adherent rodent cell lines, baby hamster kidney (BHK-21) and murine hepatoma (MH-22A), were used to examine the contribution of a cell. We tested cytotoxicity assays of the main groups of fast (non-clonogenic) methods of cell viability measuring. Plasma membrane integrity was estimated by trypan blue exclusion and LDH leakage, metabolic activity was tested by [3H]-thymidine incorporation and MTT assay, loss of monolayer adherence was measured by staining with crystal violet and CyQUANT. The most sensitive test in each case was the assay related to the site of the direct damage, and measurement of the loss of monolayer adherence proved to be as sensitive assay as the damage-specific one. All the assays applied, except for the LDH release, revealed a higher effect of combination of m-THPC-mediated phototreatment and doxorubicin compared to either of the single treatments.     

Mechanism of radiosensitization by porphyrins.

According to our previous data, hematoporphyrin dimethyl ether (HPde) at concentrations useful for photodynamic therapy can radiosensitize aggressive Ehrlich ascite carcinoma (EAT) to 2Gy irradiation inducing total tumour growth inhibition. The aim of this study was to further investigate the possible mechanism of radiosensitization of EAT by dicarboxylic porphyrin-HPde. Our results reveal that HPde is inducing several rearrangements in the EAT cells: 1.2 x 10-6 M of the photosensitizer diminishes the number of cells in mitosis by a factor of 3, increases the number of cells in the S phase of the cell cycle, modifies the activities of antioxidant enzymes glutation S-transferase (GST) and DT-diaphorase (DTD), and eventually induces slight apoptosis. Moreover, it was shown that HPde is a ligand of peripheral benzodiazepine receptor (PBR). Named "house keeper," PBR is usually responsible for all these perturbations, which, in our case, act in concert with the following ionizing radiation, producing the interaction of two antiproliferative/destructive factors.