Quantifying Oxidation of Cellulose-Associated Glucuronoxylan by Two Lytic Polysaccharide Monooxygenases from Neurospora crassa

Family AA9 lytic polysaccharide monooxygenases (LPMOs) are abundant in fungi, where they catalyze oxidative depolymerization of recalcitrant plant biomass. These AA9 LPMOs cleave cellulose and some also act on hemicelluloses, primarily other (substituted) beta-(1 -> 4)-glucans. Oxidative cleavage of xylan has been shown for only a few AA9 LPMOs, and it remains unclear whether this activity is a minor side reaction or primary function. Here, we show that Neurospora crassa LPMO9F (NcLPMO9F) and the phylogenetically related, hitherto uncharacterized NcLPMO9L from N. crassa are active on both cellulose and cellulose-associated glucuronoxylan but not on glucuronoxylan alone. A newly developed method for simultaneous quantification of xylan-derived and cellulose-derived oxidized products showed that NcLPMO9F preferentially cleaves xylan when acting on a cellulosebeechwood glucuronoxylan mixture, yielding about three times more xylan-derived than cellulose-derived oxidized products. Interestingly, under similar conditions, NcLPMO9L and the previously characterized McLPMO9H, from Malbranchea cinnamomea, showed different xylan-to-cellulose preferences, giving oxidized product ratios of about 0.5:1 and 1:1, respectively, indicative of functional variation among xylanactive LPMOs. Phylogenetic and structural analysis of xylan-active AA9 LPMOs led to the identification of characteristic structural features, including unique features that do not occur in phylogenetically remote AA9 LPMOs, such as four AA9 LPMOs whose lack of activity toward glucuronoxylan was demonstrated in the present study. Taken together, the results provide a path toward discovery of additional xylanactive LPMOs and show that the huge family of AA9 LPMOs has members that preferentially act on xylan. These findings shed new light on the biological role and industrial potential of these fascinating enzymes. IMPORTANCE Plant cell wall polysaccharides are highly resilient to depolymerization by hydrolytic enzymes, partly due to cellulose chains being tightly packed in microfibrils that are covered by hemicelluloses. Lytic polysaccharide monooxygenases (LPMOs) seem well suited to attack these resilient copolymeric structures, but the occurrence and importance of hemicellulolytic activity among LPMOs remain unclear. Here, we show that certain AA9 LPMOs preferentially cleave xylan when acting on a cellulose-glucuronoxylan mixture, and that this ability is the result of protein evolution that has resulted in a clade of AA9 LPMOs with specific structural features. Our findings strengthen the notion that the vast arsenal of AA9 LPMOs in certain fungal species provides functional versatility and that AA9 LPMOs may have evolved to promote oxidative depolymerization of a wide variety of recalcitrant, copolymeric plant polysaccharide structures. These findings have implications for understanding the biological roles and industrial potential of LPMOs

Fluoride content and recharge ability of five glassionomer dental materials

<p>Abstract</p> <p>Background</p> <p>The relationship between fluoride content and fluoride release for glass-ionomer cements is not well understood. The aim of this laboratory study was: to determine the fluoride concentrations at the surfaces of glass-ionomer materials with respect to different storage media and different pH environments; to examine the recharge ability of the materials after NaF immersion; and to assess the morphological changes at the material surfaces using scanning electron microscope and energy dispersive spectroscopic techniques (SEM/EDS).</p> <p>Methods</p> <p>Five glass-ionomer materials, Fuji Triage (FT), Fuji II LC (FII), Fuji VIII (FVIII), Fuji IX GP (FIX), and Ketac N100 (KN), were analyzed in this study. Resin-based fluoride releasing material Helioseal F (HSF) was used as a comparison material. The sample consisted of 120 cured cement disks (n = 20 disks of each tested material, 10 × 1.5 mm). Five disks of each material were stored in 4 different storage media (I- saline, II- acidic solution ph = 2.5, III- acid solution ph = 5.5, IV- NaF solution (c = 500/106). After 7 days, two disks of each material were transferred from media I, II and III to the NaF solution for 3 min. EDS analysis was conducted in 3 randomly selected spots of each experimental disk. SEM was used to determine morphological characteristics of the material surface. Differences between the experimental groups have been analyzed using Student's t-test with the level of significance set at p < 0.001.</p> <p>Results</p> <p>FT showed the highest fluoride content at the surface of the material. The lowest amounts of fluoride ions were detected at the surfaces of the FT disks stored at low pH environments, and this difference was statistically significant (p < 0.001). Glass-ionomers showed significantly higher fluoride concentrations when compared to the HSF (p < 0.001). After immersion in the NaF solution, fluoride concentrations at the surfaces of the disks increased when compared with previous storage media (FT>FVIII>KN>FII>FIX). SEM analysis of the surface morphology revealed numerous voids, cracks and microporosities in all experimental groups, except for KN and HSF. More homogenous material structure with more discrete cracks was observed in samples stored at neutral pH environment, compared to disks stored in acidic solutions.</p> <p>Conclusion</p> <p>The tested materials could be considered as promising dental materials with potential prophylactic characteristics due to their relatively high fluoride content, but also the ability to extensively reabsorb fluoride ions, especially in acidic environments.</p

Global wealth disparities drive adherence to COVID-safe pathways in head and neck cancer surgery

Peer reviewe

Management of anaphylaxis due to COVID-19 vaccines in the elderly

Older adults, especially men and/or those with diabetes, hypertension, and/or obesity, are prone to severe COVID-19. In some countries, older adults, particularly those residing in nursing homes, have been prioritized to receive COVID-19 vaccines due to high risk of death. In very rare instances, the COVID-19 vaccines can induce anaphylaxis, and the management of anaphylaxis in older people should be considered carefully. An ARIA-EAACI-EuGMS (Allergic Rhinitis and its Impact on Asthma, European Academy of Allergy and Clinical Immunology, and European Geriatric Medicine Society) Working Group has proposed some recommendations for older adults receiving the COVID-19 vaccines. Anaphylaxis to COVID-19 vaccines is extremely rare (from 1 per 100,000 to 5 per million injections). Symptoms are similar in younger and older adults but they tend to be more severe in the older patients. Adrenaline is the mainstay treatment and should be readily available. A flowchart is proposed to manage anaphylaxis in the older patients.Peer reviewe

Gastrointestinal non-infectious complications in patients on peritoneal dialysis

© 2016, University of Kragujevac, Faculty of Science, All rights reserved. Gastrointestinal complications are common among patients on peritoneal dialysis. Risk factors for the development of gastrointestinal complications in this patient population include: toxic effects of uremic toxins, frequent use of nonsteroidal anti-inflammatory drugs, Helicobacter pylori infection, angiodysplasia, increased intra-abdominal pressure, use of bioincompatible solution for peritoneal dialysis, increased glucose in solutions for peritoneal dialysis, secondary hyperparathyroidism (hypercalcemia), a disorder of lipid metabolism (hypertriglyceridemia), and the duration of peritoneal dialysis treatment. The most important non-infectious gastrointestinal complications in patients on peritoneal dialysis are: gastrointestinal bleeding, herniation and leaking of the dialysate from the abdomen (increased intra-abdominal pressure), impaired lung function (intra-abdominal hypertension), acute pancreatitis, and encapsulating sclerosis of the peritoneum. Intra-abdominal hypertension is defined as IAP ≥ 12 mmHg. Pouring the peritoneal dialysis solution leads to increased intra-abdominal pressure, which results in the development of hernias, pleuro-peritoneal dialysate leakage (hydrothorax), and restrictive pulmonary dysfunction. Risk factors for the development of acute pancreatitis in this patient population include: uraemia, secondary hyperparathyroidism with hypercalcemia, hypertriglyceridemia, features of the peritoneal dialysis solution (osmolarity, acidity, glucose, chemical irritation, and calcium in the solution for peritoneal dialysis lead to “local hypercalcemia”), toxic substances from the dialysate, the bags and tubing, and peritonitis and treatment of peritonitis with antibiotics and anticoagulants. Encapsulating sclerosis of the peritoneum is rare and is the most serious complication of long-term peritoneal dialysis. It is characterized by thickening of the peritoneum, including cancer, and signs and symptoms of obstructive ileus. Diagnosis is based on clinical, laboratory and radiological parameters. Encapsulating sclerosis of the peritoneum can be indicated by an AR-CA-125 concentration of less than 33 U/min and a concentration of AR-IL-6 greater than 350 pg/min in the effluent of patients with ultrafiltration weakness. Treatment consists of stopping peritoneal dialysis, using anti-inflammatory (corticosteroids) and anticicatricial drugs (tamoxifen), while surgical treatment includes enterolysis and adhesiolysis

The complement system and glomerular kidney diseases: Etiopathogenesis, diagnostics and treatment

© 2015, Serbian Medical Society. All rights reserved. The human immune system consists of the innate and the acquired immunity. The complement system is part of the innate immune response and it can be activated through one of the three pathways: the classical pathway, the alternative pathway and the lectin pathway. Complement activation forms the C3-convertase complex. The C3-convertase proteolytically cleaves the C3 component into C3a and C3b fragments. The C3b fragment binds to the C3-convertase to form the C5-convertase. The C5-convertase is an enzyme complex that cleaves the C5 component into C5a and C5b fragments. The C5b component binds to C6, C7, C8, C9 thus forming the terminal complex of the complement activation-the membrane attack complex (MAC) which is responsible for effector functions of the complement system. The serum complement is a mediator of C3 glomerulopathy (C3G), a group of diseases that includes membranoproliferative glomerulonephritis type II (MPGN II) or dense deposit disease (DDD) and C3 glomerulonephritis (C3GN). These diseases are caused by a dysregulation of the alternative pathway of the complement activation system. The C3 nephritic factor (C3NeF) is an autoantibody to the alternative pathway C3-convertase, which can cause dysregulation of the pathway. Newly acquired knowledge has enabled therapeutic applications of a specific antibody to the C5 complement component (eculizumab). Eculizumab is a recombinant monoclonal antibody that exhibits high affinity for the complement component C5, with the aim to inhibit its cleavage and activation of the terminal complement pathway

Secondary hypertension: Differential diagnosis and basic principles of treatment

© 2016, University of Kragujevac, Faculty of Science. All rights reserved. Secondary hypertension occurs in 5-10% of cases in the patient population with primary hypertension. e most common forms of secondary hypertension are as follows: parenchymal renal disease (renoparenchymal hypertension), renal artery stenosis (renovascular hypertension), adrenal gland adenoma (primary hyperaldosteronism), a tumour of the adrenal gland marrow (pheochromocytoma) and adenoma of adrenal and pituitary glands (Cushing’s syndrome). In patients with a typical clinical picture of secondary hypertension, the appropriate diagnostic tests should be conducted based on the suspected form of secondary hypertension. Determining a diagnosis of secondary hypertension is gradual. First, the appropriate screening tests are performed. If the screening test is positive, then additional tests to confirm the forms of secondary hypertension are conducted. Once a diagnosis of the appropriate form of secondary hypertension is confirmed, tests to distinguish causes and laterality tests to determine the precise localisation of the pathological process are applied to evaluate the response to therapy. Analysing the results of endocrine diagnostic tests provides an accurate diagnosis and selection of optimal therapeutic procedures

Beta-2 microglobulin removal with postdilution online hemodiafiltration – comparison of three different dialysis membranes

Introduction Accumulation of middle molecular weight uremic toxins causes various complications in chronic hemodialysis (HD) patients. Postdilution online hemodiafiltration (OL-HDF) efficiently removes these molecules. This study aimed to assess the effectiveness of three different dialysis membranes in removing β2-microglobulin (β2 m) within a single session of postdilution OL-HDF. Method A prospective single-center study was carried out in 30 patients (23 males and seven females, average age 54.87 ± 11.66 years, time on dialysis 4.95 ± 5.4 years) on maintenance HD. Each patient was followed for three consecutive weeks on OL-HDF with three different dialyzers: DiacapPro 19H, FX CorDiax 800, and Elisio 21H, randomly switched weekly. The reduction ratios (RR) of β2 m and albumin were compared individually. The results were analyzed with the Kolmogorov–Smirnov test, ANOVA, and the Kruskal–Wallis test. Results The average convective volume for all patients was 21.38 ± 2.97 L/session. β2-m RR was 70.86 ± 6.87%, 74.69 ± 6.51%, and 70.04 ± 9.37% with Diacap Pro 19H, FX CorDiax 800 and Elisio 21H membrane, respectively (p = 0.054). Albumin RR was 6.20 ± 2.12% with Diacap Pro 19H membrane, 6.01 ± 2.97% with FX CorDiax 800 membrane, and 6.46 ± 2.91% with Elisio 21H membrane (p = 0.812). Albumin loss was < 4 g/dialysis treatment for all membranes. Conclusion All investigated membranes effectively remove β2-m in postdilution OL-HDF with a tolerable albumin loss. The highest β2-m RR was determined for FX CorDiax 800 membrane, but with no statistically significant difference