Superior protection conferred by inactivated whole virus vaccine over subunit and DNA vaccines against salmonid alphavirus infection in Atlantic salmon (Salmo salar L.)

Salmonid alphavirus 3 (SAV-3) is an emerging pathogen in Norwegian salmon farming and causes severe annual losses. We studied the immunogenicity and protective ability of subunit and DNA vaccines based on E1 and E2 spike proteins of salmonid alphavirus subtype 3 (SAV-3), and compared these to an experimental inactivated, whole virus (IWV) vaccine in Atlantic salmon. The antigens were delivered as water-in-oil emulsions for the subunit and inactivated vaccines and non-formulated for the DNA vaccines. The IWV and the E2 subunit prime-boost groups had circulating neutralizing antibodies at challenge, correlating with high protection against lethal challenge and 3-log(10) reduction of virus titer in heart for the IWV group. Prime-boost with E1 subunit vaccine also conferred significant protection against mortality, but did not correlate with neutralizing antibody levels. Protection against pathology in internal organs was only seen for the IWV group. Prime-boost with E1 and E2 DNA vaccines showed marginal protection in terms of reduction of viral replication in target organs and protection against mortality was not different from controls. The IWV group showed significant upregulation of IFNγ and IL2 mRNA expression at 4 weeks post challenge possibly indicating that other mechanisms in addition to antibody responses play a role in mediating protection against infection. This is the first report comparing the immunogenicity and protection against mortality for IWV vaccines and spike protein subunit and DNA vaccines against salmonid alphavirus infection in Atlantic salmon. The IWV vaccine has superior immunogenicity over sub-unit and DNA vaccines.     

RAMP2 and RAMP3 mRNA levels are increased in failing rat cardiomyocytes and associated with increased responsiveness to adrenomedullin

Adrenomedullin (AM) is a potent vasorelaxing peptide with natriuretic and diuretic actions. Recent data indicate that AM may function as an endogenous regulator of cardiac function. We investigated to what extent AM, the AM receptor subtypes, and AM receptor-associated proteins were regulated in cardiomyocytes and non-cardiomyocytes of rats with congestive heart failure (CHF), and whether such regulation was paralleled by corresponding alterations of functional responses to AM. Cardiomyocytes and non-cardiomyocytes were isolated from myocardial tissue of rats 7 days after induction of myocardial infarction or sham operation. AM immunoreactivity was found in cardiomyocytes, endothelial cells, and fibroblasts. Robust increase of AM mRNA levels was observed both in the cardiomyocytes and in the non-cardiomyocytes of CHF rats compared to that of sham-operated rats (2.7-fold and 3.7-fold, respectively, P <0.05). Fairly high mRNA levels and immunoreactivity against the AM receptor chaperone receptor activity-modifying protein-2 (RAMP2) were also detected in the cardiomyocytes and non-cardiomyocytes. However, induction of RAMP2 mRNA expression was restricted to cardiomyocytes (1.8-fold increase in cardiomyocytes from CHF rats vs. sham rats; P <0.05). In contrast, very low levels of RAMP3 mRNA were observed. RAMP3 mRNA levels, however, were elevated in both cardiomyocytes and non-cardiomyocytes from CHF rats (6.5-fold and 2.4-fold increase vs. sham rats, respectively; P <0.05). Parallel increases of specific AM receptor binding sites and of AM-stimulated adenylyl cyclase activities were observed in failing cardiomyocytes compared to cardiomyocytes from sham rats (fivefold and sixfold increase, respectively; P <0.05). Thus, this study demonstrates that AM mRNA levels, AM receptor binding sites, and AM-stimulated adenylyl cyclase activities are increased in cardiomyocytes from failing rat hearts. Furthermore, our data suggest that induction of RAMP2 and RAMP3 contributes to the increased responsiveness to AM in failing cardiomyocytes.     

cDNA microarray analysis of non-selected cases of acute myeloid leukemia demonstrates distinct clustering independent of cytogenetic aberrations and consistent with morphological signs of differentiation

Acute myeloid leukemia (AML) is a heterogeneous disease with respect to biology and clinical course. Until now the basis for prognostic evaluation and therapeutic decision has been the karyotype, genetic FLT3 abnormalities and the initial chemotherapy response. A question that has emerged is if extensive gene expression analysis may supplement or partly replace current diagnostics. In an attempt to address this question, we performed cDNA microarray analysis on peripheral blood samples of 25 patients with newly diagnosed AML with high blast counts. The patients were randomly selected from a large group of consecutive patients. Leave-one-out crossvalidation (LOOCV) showed with high accuracy that gene expression classifiers could predict if leukaemia samples belonged to the FAB AML-M1 or to the FAB AML-M2 groups. An unsupervised two-dimensional hierarchical cluster analysis generated 3 patient subgroups. Except for an accumulation of samples classified as FAB M1 and M2 in cluster 3, there was no evident relationship between the clusters and the FAB classification. Each subgroup displayed clearly distinguished gene expression patterns validated using real-time quantitative PCR analysis. The identification of specific gene expressions that together constitute regulatory modules must complement cluster analyses in order to achieve an accurate basis for prognosis and prediction.     

Increased Brain Uptake of Copper and Zinc in Mice Caused by Diethyldithiocarbamate

Abstract Mice given ⁶⁴CuCl₂ and ⁶⁵ZnCl₂ (10 μmol/kg) were treated with sodium diethyldithiocarbamate (0.5 mmol/kg). The treatment increased the brain level of radioactive copper five-fold and that of radioactive zinc three-fold. Such redistribution of metal ions may be explained from the formation of lipophilic metal chelates. The increased brain levels may involve neurotoxic effects.     

Use of magnetic resonance imaging to define the anatomical location closest to all three cords of the infraclavicular brachial plexus

Infraclavicular techniques are often used to perform brachial plexus blocks. In our volunteer study we used magnetic resonance imaging to identify the brachial plexus and axillary vessels in a sagittal plane corresponding to the lateral sagittal infraclavicular block. In 20 volunteers, all cords were positioned within 2 cm from the artery approximately within 2/3 of a circle. We derived an injection site that was closest to all cords, cranio-posterior and adjacent to the axillary artery. We conclude that this knowledge may be useful for the performance of infraclavicular blocks aided by ultrasound. However, our proposals should be tested by clinical studies.     

The Effect of a Benzalkonium Chloride-Containing Nasal Spray on Human Respiratory Mucosa in vitro as a Function of Concentration and Time of Action

Abstract: Human respiratory mucosa was exposed to oxymetazoline nasal spray in varying concentrations and for varying periods of time in vitro. The drug destroyed the tissue in a concentration- and time-dependent manner. In the experiments with various concentrations of the spray, some tissue fragments retained their viability throughout the experiment. This number increased parallel to a decrease in concentrations of the test substance. All the tissue fragments exposed to undiluted nose spray underwent severe destructive alterations during the exposure period. These alterations appeared first and were most extensive in those exposed for the longest periods of time. It has previously been demonstrated that the toxic effect of oxymetazoline nasal spray in vitro is probably due to the preservative benzalkonium chloride. The apparent lack of consistency between the toxic effects of benzalkonium chloride in vitro and in vivo is discussed, with special reference to protective systems absent in vitro but present in vivo.     

Effect of ambient temperature on endurance performance while wearing cross-country skiing clothing

This study assessed the effects of exposure to cold (?14 and ?9?°C), cool (?4 and 1?°C) and moderate warm (10 and 20?°C) environments on aerobic endurance performance-related variables: maximal oxygen consumption (VO_2max), running time to exhaustion (TTE), running economy and running speed at lactate threshold (LT). Nine male endurance athletes wearing cross-country ski racing suit performed a standard running test at six ambient temperatures in a climatic chamber with a wind speed of 5?m?s^?1. The exercise protocol consisted of a 10-min warm-up period followed by four submaximal periods of 5?min at increasing intensities between 67 and 91?% of VO_2max and finally a maximal test to exhaustion. During the time course mean skin temperature decreased significantly with reduced ambient temperatures whereas T _re increased during all conditions. T _re was lower at ?14?°C than at ?9 and 20?°C. Running economy was significantly reduced in warm compared to cool environments and was also reduced at 20?°C compared to ?9?°C. Running speed at LT was significantly higher at ?4?°C than at ?9, 10 and 20?°C. TTE was significantly longer at ?4 and 1?°C than at ?14, 10 and 20?°C. No significant differences in VO_2max were found between the various ambient conditions. The optimal aerobic endurance performance wearing a cross-country ski racing suit was found to be ?4 and 1?°C, while performance was reduced under moderate warm (10 and 20?°C) and cold (?14 and ?9?°C) ambient conditions.