Multiple roles for UV RESISTANCE LOCUS8 in regulating gene expression and metabolite accumulation in arabidopsis under solar ultraviolet radiation

Photomorphogenic responses triggered by low fluence rates of ultraviolet B radiation (UV-B; 280–315 nm) are mediated by the UV-B photoreceptor UV RESISTANCE LOCUS8 (UVR8). Beyond our understanding of the molecular mechanisms of UV-B perception by UVR8, there is still limited information on how the UVR8 pathway functions under natural sunlight. Here, wild-type Arabidopsis (Arabidopsis thaliana) and the uvr8-2 mutant were used in an experiment outdoors where UV-A (315–400 nm) and UV-B irradiances were attenuated using plastic films. Gene expression, PYRIDOXINE BIOSYNTHESIS1 (PDX1) accumulation, and leaf metabolite signatures were analyzed. The results show that UVR8 is required for transcript accumulation of genes involved in UV protection, oxidative stress, hormone signal transduction, and defense against herbivores under solar UV. Under natural UV-A irradiance, UVR8 is likely to interact with UV-A/blue light signaling pathways to moderate UV-B-driven transcript and PDX1 accumulation. UVR8 both positively and negatively affects UV-A-regulated gene expression and metabolite accumulation but is required for the UV-B induction of phenolics. Moreover, UVR8-dependent UV-B acclimation during the early stages of plant development may enhance normal growth under long-term exposure to solar UV

DigiWeb - a workflow environment for quality assurance of transcription in digitization of natural history collections

Data produced by digitization increases the scientific use of natural history collections. However, in mass digitization, attention must be paid to the flawless management of the workflows, and high quantities of end results should not be compromised by a low standard of quality. A web-based environment DigiWeb was created for controlling the workflow of transcribing data from images of natural history specimens. Using DigiWeb, it was possible to manage the workflow of transcription and data proofing, include all participants to the workflow, allow collaboration and training, and also to provide useful processing features. The data emerging from this process passes quality control standards which are supported by DigiWeb and based on the strict requirements of the ISO 2859 standard

Respiratory chain complex III deficiency due to mutated BCS1L : a novel phenotype with encephalomyopathy, partially phenocopied in a Bcs1l mutant mouse model

Background: Mitochondrial diseases due to defective respiratory chain complex III (CIII) are relatively uncommon. The assembly of the eleven-subunit CIII is completed by the insertion of the Rieske iron-sulfur protein, a process for which BCS1L protein is indispensable. Mutations in the BCS1L gene constitute the most common diagnosed cause of CIII deficiency, and the phenotypic spectrum arising from mutations in this gene is wide. Results: A case of CIII deficiency was investigated in depth to assess respiratory chain function and assembly, and brain, skeletal muscle and liver histology. Exome sequencing was performed to search for the causative mutation(s). The patient's platelets and muscle mitochondria showed respiration defects and defective assembly of CIII was detected in fibroblast mitochondria. The patient was compound heterozygous for two novel mutations in BCS1L, c.306A > T and c.399delA. In the cerebral cortex a specific pattern of astrogliosis and widespread loss of microglia was observed. Further analysis showed loss of Kupffer cells in the liver. These changes were not found in infants suffering from GRACILE syndrome, the most severe BCS1L-related disorder causing early postnatal mortality, but were partially corroborated in a knock-in mouse model of BCS1L deficiency. Conclusions: We describe two novel compound heterozygous mutations in BCS1L causing CIII deficiency. The pathogenicity of one of the mutations was unexpected and points to the importance of combining next generation sequencing with a biochemical approach when investigating these patients. We further show novel manifestations in brain, skeletal muscle and liver, including abnormality in specialized resident macrophages (microglia and Kupffer cells). These novel phenotypes forward our understanding of CIII deficiencies caused by BCS1L mutations.Peer reviewe

Photoreceptors UVR8 and phytochrome B cooperate to optimize plant growth and defense in patchy canopies

Light is a critical source of information for plants. Plants use the phytochromes (particularly phyB) to detect light signals associated with the proximity of competitors. A low ratio of red (R) to far-red (FR) radiation (R:FR) indicates increased competition intensity, and triggers morphological responses that allow the plant to escape shading from its neighbors (the shade avoidance syndrome, SAS). Recent evidence from studies on light regulation of plant immunity has suggested that plants may also use ultraviolet-B (UV-B, 290-315 nm) radiation as an indicator of competition intensity and light availability. In addition, recent studies have shown that UV-B radiation can strongly repress SAS responses triggered by low R:FR ratios. Ambient UV-B radiation causes damaging effects on plants, such as DNA damage, and also induces adaptive photomorphogenic responses acting through a specific UV-B photoreceptor (UVR8). Therefore, the possibility exists that plants integrate information perceived by phyB and UVR8 to make decisions about growth and defense when faced with a complex light environment, such as the one that characterizes vegetation canopies. In this Letter, we address this possibility and discuss how the interplay between UV-B and R:FR signaling fine tunes plant growth and defense to optimize resource utilization in patchy canopy environments.Fil: Mazza, Carlos Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; ArgentinaFil: Ballare, Carlos Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico Chascomús. Instituto de Investigaciones Biotecnológicas (sede Chascomús); Argentin

Brain inflammation is accompanied by peripheral inflammation in Cstb(-/-) mice, a model for progressive myoclonus epilepsy

Progressive myoclonus epilepsy of Unverricht-Lundborg type (EPM1) is an autosomal recessively inherited childhood-onset neurodegenerative disorder, characterized by myoclonus, seizures, and ataxia. Mutations in the cystatin B gene (CSTB) underlie EPM1. The CSTB-deficient (Cstb(-/-)) mouse model recapitulates key features of EPM1, including myoclonic seizures. The mice show early microglial activation that precedes seizure onset and neuronal loss and leads to neuroinflammation. We here characterized the inflammatory phenotype of Cstb(-/-) mice in more detail. We found higher concentrations of chemokines and pro-inflammatory cytokines in the serum of Cstb(-/-) mice and higher CXCL13 expression in activated microglia in Cstb(-/-) compared to control mouse brains. The elevated chemokine levels were not accompanied by blood-brain barrier disruption, despite increased brain vascularization. Macrophages in the spleen and brain of Cstb(-/-) mice were predominantly pro-inflammatory. Taken together, these data show that CXCL13 expression is a hallmark of microglial activation in Cstb(-/-)mice and that the brain inflammation is linked to peripheral inflammatory changes, which might contribute to the disease pathology of EPM1.Peer reviewe

Gene-Expression Profiling Suggests Impaired Signaling via the Interferon Pathway in Cstb(-/-) Microglia

Progressive myoclonus epilepsy of Unverricht-Lundborg type (EPM1, OMIM254800) is an autosomal recessive neurodegenerative disorder characterized by stimulus-sensitive and action-activated myoclonus, tonic-clonic epileptic seizures, and ataxia. Loss-of-function mutations in the gene encoding the cysteine protease inhibitor cystatin B (CSTB) underlie EPM1. The deficiency of CSTB in mice (Cstb(-/-) mice) generates a phenotype resembling the symptoms of EPM1 patients and is accompanied by microglial activation at two weeks of age and an upregulation of immune system-associated genes in the cerebellum at one month of age. To shed light on molecular pathways and processes linked to CSTB deficiency in microglia we characterized the transcriptome of cultured Cstb(-/-) mouse microglia using microarray hybridization and RNA sequencing (RNA-seq). The gene expression profiles obtained with these two techniques were in good accordance and not polarized to either pro- or anti-inflammatory status. In Cstb(-/-) microglia, altogether 184 genes were differentially expressed. Of these, 33 genes were identified by both methods. Several interferon-regulated genes were weaker expressed in Cstb(-/-) microglia compared to control. This was confirmed by quantitative real-time PCR of the transcripts Irf7 and Stat1. Subsequently, we explored the biological context of CSTB deficiency in microglia more deeply by functional enrichment and canonical pathway analysis. This uncovered a potential role for CSTB in chemotaxis, antigen-presentation, and in immune-and defense response-associated processes by altering JAK-STAT pathway signaling. These data support and expand the previously suggested involvement of inflammatory processes to the disease pathogenesis of EPM1 and connect CSTB deficiency in microglia to altered expression of interferon-regulated genes.Peer reviewe

Proteomic mapping of differentially vulnerable pre-synaptic populations identifies regulators of neuronal stability in vivo.

Synapses are an early pathological target in many neurodegenerative diseases ranging from well-known adult onset conditions such as Alzheimer and Parkinson disease to neurodegenerative conditions of childhood such as spinal muscular atrophy (SMA) and neuronal ceroid lipofuscinosis (NCLs). However, the reasons why synapses are particularly vulnerable to such a broad range of neurodegeneration inducing stimuli remains unknown. To identify molecular modulators of synaptic stability and degeneration, we have used the Cln3-/- 33 mouse model of a juvenile form of NCL. We profiled and compared the molecular composition of anatomically-distinct, differentially-affected pre-synaptic populations from the Cln3-/- 35 mouse brain using proteomics followed by bioinformatic analyses. Identified protein candidates were then tested using a Drosophila CLN3 model to study their ability to modify the CLN3-neurodegenerative phenotype in vivo. We identified differential perturbations in a range of molecular cascades correlating with synaptic vulnerability, including valine catabolism and rho signalling pathways. Genetic and pharmacological targeting of key ‘hub’ proteins in such pathways was sufficient to modulate phenotypic presentation in a Drosophila CLN3 model. We propose that such a workflow provides a target rich method for the identification of novel disease regulators which could be applicable to the study of other conditions where appropriate models exist

Effects on blood pressure after treatment of obstructive sleep apnea with an oral appliance with mandibular advancement, a 3 year follow-up

The study purpose was to investigate if rduction of obstructive sleep apnea with oral appliance therapy (AO)affects blood pressure (BP) in hypertensive patients in a short-term and long-term perspective. Material and methods: Twenty-nine consecutive patients with verified obstructive sleep apnea (OSA);defined as apnea index (AI) <5 and/or panea-hypoponea index (AHI) <10/hours, received as a treatment an OA with mandibular advancement. The reference BP was measured at 3 study visits; before treatment, after 3 months and after 3 yrs of treatment, respectively. The CP was measured twice at those visits with an automatic unit and the second value was registered as the CP of the visit. The treatment effect of OSA was measured after 3 months use by are repeated somnographic registration wearing the OA. At this short-term evaluation after 3 months the patients were asked about the compliance and the frequency of OA use nights/week. At the long-term follow up after 3 years use of OA the patients answered questions about altering in and hypotensive medication, frequency in use of OA (nights/week), weight gain or loss and a self-report of general well-being. A complete response to treatment was considered as success when a normalization of nightly breathing occurred, defined as AHI<10. Results: Between baseline and the short-term evaluation 4 patients changed treatment. A complete treatment response of OSA was achieved in 25 of 29 patients. The remaining 25 patients were called for a long-term follow-up after 3 years. Three patients were unable to participate. At the 3 years follow-up the remaining 22 treated patients had a statistically significant reduction for the sustolic BP of -15.4mm Hg and for the diastolic BP of -10.3mm Hg. This significance was received between baseline and the short-term evaluation (p<0.001), and remained at the long-term follow-up. All 22 patients reported a good general well being after 3 years of treatment. Conclusion: OA therapy reduced blood pressure in both short-term and long-term perspectives in patients with OSA and hypertension