Inhibition of rubella virus replication by the broad-spectrum drug nitazoxanide in cell culture and in a patient with a primary immune deficiency

Persistent rubella virus (RV) infection has been associated with various pathologies such as congenital rubella syndrome, Fuchs's uveitis, and cutaneous granulomas in patients with primary immune deficiencies (PID). Currently there are no drugs to treat RV infections. Nitazoxanide (NTZ) is an FDA-approved drug for parasitic infections, and has been recently shown to have broad-spectrum antiviral activities. Here we found that empiric 2-month therapy with oral NTZ was associated in the decline/elimination of RV antigen from lesions in a PID patient with RV positive granulomas, while peginterferon treatment had no effect. In addition, we characterized the effects of NTZ on cell culture models of persistent RV infection. NTZ significantly inhibited RV replication in a primary culture of human umbilical vein endothelial cells (HUVEC) and Vero and A549 epithelial cell lines in a dose dependent manner with an average 50% inhibitory concentration of 0.35 mu g/ml (1.1 mu M). RV strains representing currently circulating genotypes were inhibited to a similar extent. NTZ affected early and late stages of infection by inhibiting synthesis of cellular and RV RNA and interfering with intracellular trafficking of the RV surface glycoproteins, E1 and E2. These results suggest a potential application of NTZ for the treatment of persistent rubella infections, but more studies are required.Peer reviewe

Environment simulator for studying automatic crop farming

Agricultural machines capable of utilizing variable rate application technology are tackling spatial variability in agricultural fields.  Agricultural field robots are the next step in technology, robots which are capable of utilizing sensor and actuating technologies without human contact and operate only areas of interest.  However, agricultural field robots are still under research.  Robots are just one part of the next generation of crop farming having more advanced tools to do the work which currently requires humans.  The next generation of crop farming, in the vision of the authors, is based on automation, which incorporates stationary and moving sensors systems, robots, model based decision making, automated operation planning which adapts to spatial variability according to the measurements as well as to weather conditions.  This article presents a top-down approach of automated crop farming using simulation, trying to cover all the component parts on a fully automated farm.  In the article, the developed simulation platform is presented as well as sample simulation results.  The environment simulator is based on crop growth models, weed growth models, soil models, spatial variation generation and weather statistics.  Models for the environment were found in literature and were tailored and tuned to fit the simulation purposes, to form a collection of models.  The collection of models was evaluated by using sensitivity analysis.  Furthermore, a full scale scenario was simulated over one season, incorporating 9000 spatial cells in five fields of a farm.   Keywords: robots, crop growth models, soil water models, decision making, operation plannin

Effect of barley grinding method and sodium polyacrylate supplement in the diet on the performance and stomach ulcer development of growing finishing pigs

Two different grinding methods - rolling and hammer milling - as well as polyacrylate supplement in the diet were studied to evaluate their effect on the performance of pigs and the incidence of gastric lesions. The experiment was carried out in 2 x 2 factorial arrangement with a total of 160 pigs. The grist size of rolled barley was bigger than of hammermilled barley, but the difference in water-binding capacity was insignificant. No significant differences were observed in the performance traits of pigs fed either rolled or hammer-milled barley. The sodium polyacrylate supplement had no effect on the daily gain, feed:gain ratio or carcass quality of the pigs. Gastric ulcers and constrictions of the oesophageal opening of the stomach were more frequent in the groups fed hammer-milled barley than in the groups fed rolled barley, the difference being statistically significant (

Adenoviral Gene Transfer Restores Lysyl Hydroxylase Activity in Type VI Ehlers-Danlos Syndrome

Type VI Ehlers-Danlos syndrome is a disease characterized by disturbed lysine hydroxylation of collagen. The disease is caused by mutations in lysyl hydroxylase 1 gene and it affects several organs including the cardiovascular system, the joint and musculoskeletal system, and the skin. The skin of type VI Ehlers-Danlos syndrome patients is hyperelastic, scars easily, and heals slowly and poorly. We hypothesized that providing functional lysyl hydroxylase 1 gene to the fibroblasts in and around wounds in these patients would improve healing. In this study we tested the feasibility of transfer of the lysyl hydroxylase 1 gene into fibroblasts derived from rats and a type VI Ehlers-Danlos syndrome patient (in vitro) and into rat skin (in vivo). We first cloned human lysyl hydroxylase 1 cDNA into a recombinant adenoviral vector (Ad5RSV-LH). Transfection of human type VI Ehlers-Danlos syndrome fibroblasts (about 20% of normal lysyl hydroxylase 1 activity) with the vector increased lysyl hydroxylase 1 activity in these cells to near or greater levels than that of wild type, unaffected fibroblasts. The adenoviral vector successfully transfected rat fibroblasts producing both β-galactosidase and lysyl hydroxylase 1 gene activity. We next expanded our studies to a rodent model. Intradermal injections of the vector to the abdominal skin of rats produced lysyl hydroxylase 1 mRNA and elevated lysyl hydroxylase 1 activity, in vivo. These data suggest the feasibility of gene replacement therapy to modify skin wound healing in type VI Ehlers-Danlos syndrome patients

Self-Rated Mental Stress and Exercise Training Response in Healthy Subjects

Purpose: Individual responses to aerobic training vary from almost none to a 40% increase in aerobic fitness in healthy subjects. We hypothesized that the baseline self-rated mental stress may influence to the training response. Methods: The study population included 44 healthy sedentary subjects (22 women) and 14 controls. The laboratory controlled training period was 2 weeks, including five sessions a week at an intensity of 75% of the maximum heart rate for 40 min/session. Self-rated mental stress was assessed by inquiry prior to the training period from 1 (low psychological resources and a lot of stressors in my life) to 10 (high psychological resources and no stressors in my life), respectively. Results: Mean peak oxygen uptake (VO2peak) increased from 34 ± 7 to 37 ± 7 ml kg−1 min−1 in training group (p < 0.001) and did not change in control group (from 34 ± 7 to 34 ± 7 ml kg−1 min−1). Among the training group, the self-rated stress at the baseline condition correlated with the change in fitness after training intervention, e.g., with the change in maximal power (r = 0.45, p = 0.002, W/kg) and with the change in VO2peak (r = 0.32, p = 0.039, ml kg−1 min−1). The self-rated stress at the baseline correlated with the change in fitness in both female and male, e.g., r = 0.44, p = 0.039 and r = 0.43, p = 0.045 for ΔW/kg in female and male, respectively. Conclusion: As a novel finding the baseline self-rated mental stress is associated with the individual training response among healthy females and males after highly controlled aerobic training intervention. The changes in fitness were very low or absent in the subjects who experience their psychological resources low and a lot of stressors in their life at the beginning of aerobic training intervention

Elevated Cerebrospinal Fluid Neopterin Concentration Is Associated with Disease Severity in Acute Puumala Hantavirus Infection

Peer reviewe

Effects of bright light treatment on psychomotor speed in athletes

Purpose: A recent study suggests that transcranial brain targeted light treatment via ear canals may have physiological effects on brain function studied by functional magnetic resonance imaging (fMRI) techniques in humans. We tested the hypothesis that bright light treatment could improve psychomotor speed in professional ice hockey players. Methods: Psychomotor speed tests with audio and visual warning signals were administered to a Finnish National Ice Hockey League team before and after 24 days of transcranial bright light or sham treatment. The treatments were given during seasonal darkness in the Oulu region (latitude 65 degrees north) when the strain on the players was also very high (10 matches during 24 days). A daily 12-min dose of bright light or sham (n = 11 for both) treatment was given every morning between 8–12 am at home with a transcranial bright light device. Mean reaction time and motor time were analyzed separately for both psychomotor tests. Analysis of variance for repeated measures adjusted for age was performed. Results: Time x group interaction for motor time with a visual warning signal was p = 0.024 after adjustment for age. In Bonferroni post-hoc analysis, motor time with a visual warning signal decreased in the bright light treatment group from 127 ± 43 to 94 ± 26 ms (p = 0.024) but did not change significantly in the sham group 121 ± 23 vs. 110 ± 32 ms (p = 0.308). Reaction time with a visual signal did not change in either group. Reaction or motor time with an audio warning signal did not change in either the treatment or sham group. Conclusion: Psychomotor speed, particularly motor time with a visual warning signal, improves after transcranial bright light treatment in professional ice-hockey players during the competition season in the dark time of the year

Haploinsufficiency of A20 impairs protein–protein interactome and leads into caspase-8-dependent enhancement of NLRP3 inflammasome activation

Objectives TNFAIP3 encodes A20 that negatively regulates nuclear factor kappa light chain enhancer of activated B cells (NF-κB), the major transcription factor coordinating inflammatory gene expression. TNFAIP3 polymorphisms have been linked with a spectrum of inflammatory and autoimmune diseases and, recently, loss-of-function mutations in A20 were found to cause a novel inflammatory disease ‘haploinsufficiency of A20’ (HA20). Here we describe a family with HA20 caused by a novel TNFAIP3 loss-of-function mutation and elucidate the upstream molecular mechanisms linking HA20 to dysregulation of NF-κB and the related inflammasome pathway.Methods NF-κB activation was studied in a mutation-expressing cell line using luciferase reporter assay. Physical and close-proximity protein–protein interactions of wild-type and TNFAIP3 p.(Lys91*) mutant A20 were analysed using mass spectrometry. NF-κB -dependent transcription, cytokine secretion and inflammasome activation were compared in immune cells of the HA20 patients and control subjects.Results The protein–protein interactome of p.(Lys91*) mutant A20 was severely impaired, including interactions with proteins regulating NF-κB activation, DNA repair responses and the NLR family pyrin domain containing 3 (NLRP3) inflammasome. The p.(Lys91*) mutant A20 failed to suppress NF-κB signalling, which led to increased NF-κB -dependent proinflammatory cytokine transcription. Functional experiments in the HA20 patients’ immune cells uncovered a novel caspase-8-dependent mechanism of NLRP3 inflammasome hyperresponsiveness that mediated the excessive secretion of interleukin-1β and interleukin-18.Conclusions The current findings significantly deepen our understanding of the molecular mechanisms underlying HA20 and other diseases associated with reduced A20 expression or function, paving the way for future therapeutic targeting of the pathway.Peer reviewe