Recombinant production and regioselectivity study of lytic polysaccharide monooxygenases

Our dependence of the oil industry is one of today’s major concerns worldwide. Plant biomass has the potential to replace a large fraction of fossil resources for the production of fuels, chemicals and materials. Lignocellulose is, however, very recalcitrant towards enzymatic degradation, representing one of the biggest hurdles to cost-effective bio-refining. In this regard, a major breakthrough came with the discovery of lytic polysaccharide monooxygenases (LPMOs). These enzymes oxidatively cleave and disrupt the ordered structure of cellulose, making it better accessible for classical cellulases. Therefore, these enzymes are currently attracting much scientific attention. Although they all have a relatively similar active site, the target of their oxidative activity is known to differ amongst LPMOs. In this thesis, an extensive mutagenesis study was performed to better understand the determinants of this oxidative regioselectivity. For this, four fungal and two bacterial LPMO representatives were produced in Pichia pastoris and Escherichia coli, respectively. Carefully studying the mixture of products released from cellulose by these enzymes and their mutants, showed that a complex matrix of interactions on the enzyme’s substrate binding surface is not only responsible for binding cellulose, but also for accurately directing the oxidative force towards the C1 and/or C4 carbon

Is traumatic and non-traumatic neck pain associated with brain alterations? : a systematic review

Background: Chronic neck pain affects 50% - 85% of people who have experienced an acute episode. This transition and the persistence of chronic complaints are believed to be mediated by brain alterations among different central mechanisms. Objectives: This study aimed to systematically review and critically appraise the current existing evidence regarding structural and functional brain alterations in patients with whiplash associated disorders (WAD) and idiopathic neck pain (INP). Additionally, associations between brain alterations and clinical symptoms reported in neck pain patients were evaluated. Study Design: Systematic review. Methods: The present systematic review was performed according to the PRISMA guidelines. PubMed, Web of Science, and Cochrane databases were searched. First, the obtained articles were screened based on title and abstract. Secondly, the screening was based on the full text. Risk of bias in included studies was investigated. Results: Twelve studies met the inclusion criteria. Alterations in brain morphology and function, including perfusion, neurotransmission, and blood oxygenation level dependent-signal, were demonstrated in chronic neck pain patients. There is some to moderate evidence for both structural and functional brain alterations in patients with chronic neck pain. In contrast, no evidence for structural brain alterations in acute neck pain patients was found. Limitations: Only 12 articles were included, which allows only cautious conclusions to be drawn. Conclusion: Brain alterations were observed in both patients with chronic WAD and chronic INP. Furthermore, more evidence exists for brain alterations in chronic WAD, and different underlying mechanisms might be present in both pathologies. In addition, pain and disability were correlated with the observed brain alterations. Accordingly, morphological and functional brain alterations should be further investigated in patients with chronic WAD and chronic INP with newer and more sensitive techniques, and associative clinical measurements seem indispensable in future research

The impact of different lenses on visual and musculoskeletal complaints in VDU workers with work-related neck complaints : a randomized controlled trial

Objectives: The impact of wearing lenses on visual and musculoskeletal complaints in VDU workers is currently unknown. The goal of this study was 1) to evaluate the impact of wearing VDU lenses on visual fatigue and self-reported neck pain and disability, compared to progressive lenses, and 2) to measure the effect of both lenses on head inclination and pressure pain thresholds during the performance of a VDU task. Methods: Thirty-five eligible subjects were randomly assigned to wear progressive VDU lenses (VDU group) (n = 18) or progressive lenses (P group) (n = 17). They were enquired about visual complaints (VFQ), self-perceived pain (NRS) and disability (NDI) at baseline (with old lenses), and 1 week, 3 months and 6 months after wearing their new lenses. In addition, Forward Head Angle (FHA) and PPTs were assessed during and after a VDU task before and 6 months after wearing the new lenses. A short questionnaire concerning the satisfaction about the study lenses was completed at the end of the study. Results: In both groups, visual fatigue and neck pain was decreased at 3 and 6 months follow up, compared to baseline. All PPTs were higher during the second VDU task, independent of the type of lenses. The VDU group reported a significantly higher suitability of the lenses for VDU work. Conclusion: It can be concluded that there is little difference in effect of the different lenses on visual and musculoskeletal comfort. Lenses should be adjusted to the task-specific needs and habits of the participant

A quantitative indicator diagram for lytic polysaccharide monooxygenases reveals the role of aromatic surface residues in HjLPMO9A regioselectivity

Lytic polysaccharide monooxygenases ( LPMOs) have changed our understanding of lignocellulosic degradation dramatically over the last years. These metalloproteins catalyze oxidative cleavage of recalcitrant polysaccharides and can act on the C1 and/or C4 position of glycosidic bonds. Structural data have led to several hypotheses, but we are still a long way from reaching complete understanding of the factors that determine their divergent regioselectivity. Site-directed mutagenesis enables the investigation of structure-function relationship in enzymes and will be of major importance in unraveling this intriguing matter. In this context, it is crucial to have an enzyme assay or screening approach with a direct correlation with the desired functionality. LPMOs render this search extra challenging due to their insoluble substrates, complex pattern of reaction products and lack of synthetic standards of most oxidized products. Here, we describe a regioselectivity indicator diagram based on the time-course of only 2 HPAEC-PAD signals. The diagram was successfully used to confirm the hypothesis that aromatic surface residues influence the C1/C4 oxidation ratio in Hypocrea jecorina LPMO9A. Consequently, the diagram should become a valuable tool in the search towards better understanding and engineering of regioselectivity in LPMOs

Effects of stress and relaxation on central pain modulation in chronic whiplash and fibromyalgia patients compared to healthy controls

Background: Compelling evidence has demonstrated that impaired central pain modulation contributes to persistent pain in patients with chronic whiplash associated disorders (WAD) and fibromyalgia (FM). However, there is limited research concerning the influence of stress and relaxation on central pain modulation in patients with chronic WAD and FM. Objectives: The present study aims to investigate the effects of acute cognitive stress and relaxation on central pain modulation in chronic WAD and FM patients compared to healthy individuals. Study Design: A randomized crossover design was employed. Setting: The present study took place at the University of Brussels, the University Hospital Brussels, and the University of Antwerp. Methods: Fifty-nine participants (16 chronic WAD patients, 21 FM, 22 pain-free controls) were enrolled and subjected to various pain measurements. Temporal summation (TS) of pain and conditioned pain modulation (CPM) were evaluated. Subsequently, participants were randomly allocated to either a group that received progressive relaxation therapy or a group that performed a battery of cognitive tests (= cognitive stressor). Afterwards, all pain measurements were repeated. One week later participant groups were switched. Results: A significant difference was found between the groups in the change in TS in response to relaxation (P = 0.008) and cognitive stress (P = 0.003). TS decreased in response to relaxation and cognitive stress in chronic WAD patients and controls. In contrast, TS increased after both interventions in FM patients. CPM efficacy decreased in all 3 groups in response to relaxation (P = 0.002) and cognitive stress (P = 0.001). Limitations: The obtained results only apply for a single session of muscle relaxation therapy and cognitive stress, whereby no conclusions can be made for effects on pain perception and modulation of chronic cognitive stress and long-term relaxation therapies. Conclusions: A single relaxation session as well as cognitive stress may have negative acute effects on pain modulation in patients with FM, while cognitive stress and relaxation did not worsen bottom-up sensitization in chronic WAD patients and healthy persons. However, endogenous pain inhibition, assessed using a CPM paradigm, worsened in chronic WAD and FM patients, as well as in healthy people following both interventions

Does conservative treatment change the brain in patients with chronic musculoskeletal pain? : a systematic review

Background: Chronic musculoskeletal pain is characterized by maladaptive central neuroplastic changes. Many observational studies have demonstrated that chronic pain states are associated with brain alterations regarding structure and/or function. Rehabilitation of patients with chronic musculoskeletal pain may include cognitive, exercise, or multimodal therapies. Objective: The current review aims to provide a constructive overview of the existing literature reporting neural correlates, based on brain magnetic resonance imaging (MRI) techniques, following conservative treatment in chronic musculoskeletal pain patients. Study Design: Systematic review of the literature. Methods: The current review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. Literature was searched from 3 databases and screened for eligibility. Methodological quality across studies was assessed with Cochrane Collaboration's tool for assessing risk of bias and quality of evidence was determined applying the Grades of Recommendation, Assessment, Development and Evaluation (GRADE) approach. Results: A total of 9 eligible studies were identified with a predominant high risk of bias. Cognitive behavioral therapy induced several structural and functional changes predominantly in prefrontal cortical regions and a shift from affective to sensory-discriminative brain activity after behavioral extinction training. Multidisciplinary treatment in pediatric complex regional pain syndrome facilitated normalization of functional connectivity of resting-state networks and the amygdala, and increased gray matter in prefrontal and specific subcortical areas. Exercise therapy led to specific for resting-state functional connectivity and a trend towards pressure-induced brain activity changes. Limitations: A very small number of studies was available, which furthermore exhibited small study samples. Moreover, only 2 of the included studies were randomized controlled trials. Conclusions: It is likely that conservative treatments may induce mainly functional and structural brain changes in prefrontal regions in patients with chronic musculoskeletal pain. Due to the relatively high risk of bias across the included studies, future studies with randomized designs are needed to confirm the current findings. In addition, more research evaluating

The effect of experimental low back pain on lumbar muscle activity in people with a history of clinical low back pain: a muscle functional MRI study

In people with a history of low back pain (LBP), structural and functional alterations have been observed at several peripheral and central levels of the sensorimotor pathway. These existing alterations might interact with the way the sensorimotor system responds to pain. We examined this assumption by evaluating the lumbar motor responses to experimental nociceptive input of 15 participants during remission of unilateral recurrent LBP. Quantitative T2 images (muscle functional MRI) were taken bilaterally of multifidus, erector spinae, and psoas at several segmental levels (L3 upper and L4 upper and lower endplate) and during several conditions: 1) at rest, 2) upon trunk-extension exercise without pain, and 3) upon trunk-extension exercise with experimental induced pain at the clinical pain-side (1.5-ml intramuscular hypertonic saline injections in erector spinae). Following experimental pain induction, muscle activity levels similarly reduced for all three muscles, on both painful and nonpainful sides, and at multiple segmental levels (P = 0.038). Pain intensity and localization from experimental LBP were similar as during recalled clinical LBP episodes. In conclusion, unilateral and unisegmental experimental LBP exerts a generalized and widespread decrease in lumbar muscle activity during remission of recurrent LBP. This muscle response is consistent with previous observed patterns in healthy people subjected to the same experimental pain paradigm. It is striking that similar inhibitory patterns in response to pain could be observed, despite the presence of preexisting alterations in the lumbar musculature during remission of recurrent LBP. These results suggest that motor output can modify along the course of recurrent LBP

Lack of evidence for central sensitization in idiopathic, non-traumatic neck pain : a systematic review

Background: Chronic neck pain is a common problem with a poorly understood pathophysiology. Often no underlying structural pathology can be found and radiological imaging findings are more related to age than to a patient's symptoms. Besides its common occurrence, chronic idiopathic neck pain is also very disabling with almost 50% of all neck pain patients showing moderate disability at long-term follow-up. Central sensitization (CS) is defined as "an amplification of neural signaling within the central nervous system that elicits pain hypersensitivity," "increased responsiveness of nociceptive neurons in the central nervous system to their normal or subthreshold afferent input," or "an augmentation of responsiveness of central neurons to input from unimodal and polymodal receptors." There is increasing evidence for involvement of CS in many chronic pain conditions. Within the area of chronic idiopathic neck pain, there is consistent evidence for the presence and clinical importance of CS in patients with traumatic neck pain, or whiplash-associated disorders. However, the majority of chronic idiopathic neck pain patients are unrelated to a traumatic injury, and hence are termed chronic idiopathic non-traumatic neck pain. When comparing whiplash with idiopathic non-traumatic neck pain, indications for different underlying mechanisms are found. Objective: The goal of this article was to review the existing scientific literature on the role of CS in patients with chronic idiopathic non-traumatic neck pain. Study Design: Systematic review. Setting: All selected studies were case control studies. Methods: A systematic search of existing, relevant literature was performed via the electronic databases Medline, Embase, Web of Science, Cinahl, PubMed, and Google Scholar. All titles and abstracts were checked to identify relevant articles. An article was considered eligible if it met following inclusion criteria: (1) participants had to be human adults (> 18 years) diagnosed with idiopathic non-traumatic chronic (present for at least 3 months) neck pain; (2) papers had to report outcomes related to CS; and (3) articles had to be full-text reports or original research (no abstracts, case-reports, reviews, meta-analysis, letters, or editorials). Results: Six articles were found eligible after screening the title, abstract and - when necessary the full text for in- and exclusion criteria. All selected studies were case-control studies. Overall, results regarding the presence of CS were divergent. While the majority of patients with chronic traumatic neck pain (i.e. whiplash) are characterized by CS, this is not the case for patients with chronic idiopathic neck pain. The available evidence suggests that CS is not a major feature of chronic idiopathic neck pain. Individual cases might have CS pain, but further work should reveal how they can be characterized. Limitations: Very few studies available. Conclusions: Literature about CS in patients with chronic idiopathic non-traumatic neck pain is rare and results from the available studies provide an inconclusive message. CS is not a characteristic feature of chronic idiopathic and non-traumatic neck pain, but can be present in some individuals of the population. In the future a subgroup with CS might be defined, but based on current knowledge it is not possible to characterize this subgroup. Such information is important in order to provide targeted treatment

Functional assessment of the cervical spine in F-16 pilots with and without neck pain

Introduction: Spinal symptoms in fighter pilots are a serious aeromedical problem. The most common neck complaints are muscular pain and strain. The aim of the current study was to determine possible differences in the cervical range of motion (CROM), neck position sense, and neck muscle strength between pilots with and without neck pain. Methods: There were 90 male F-I 6 pilots who volunteered, of which 17 had experienced bilateral neck pain. A standardized questionnaire was used to collect personal information. The maximum isometric neck flexion/extension and lateral flexion strength, the neck position sense, and the cervical range of motion were measured. Results: There were no significant differences between healthy pilots and those with neck pain concerning neck Muscle strength and neck position sense. The neck pain group had a limited CROM in the sagittal plane (130 degrees; CI: 116 degrees-144 degrees) and in the transversal plane (155 degrees; CI: 140 degrees-170 degrees) compared to the healthy pilots. Discussion: In the current study we screened for different motor skills so that deficits Could be detected and retraining programs could be implemented when necessary. According to our results, individual retraining programs might reduce neck pain and therefore a well-instructed training program to maintain a proper active CROM should be implemented. Future Studies should investigate the effectiveness of this kind of program