Bead arrays for antibody and complement profiling reveal joint contribution of antibody isotypes to C3 deposition

The development of antigen arrays has provided researchers with great tools to identify reactivities against self or foreign antigens from body fluids. Yet, these approaches mostly do not address antibody isotypes and their effector functions even though these are key points for a more detailed understanding of disease processes. Here, we present a bead array-based assay for a multiplexed determination of antigen-specific antibody levels in parallel with their properties for complement activation. We measured the deposition of C3 fragments from serum samples to reflect the degree of complement activation via all three complement activation pathways. We utilized the assay on a bead array containing native and citrullinated peptide antigens to investigate the levels of IgG, IgM and IgA autoantibodies along with their complement activating properties in serum samples of 41 rheumatoid arthritis patients and 40 controls. Our analysis revealed significantly higher IgG reactivity against the citrullinated fibrinogen β and filaggrin peptides as well as an IgA reactivity that was exclusive for citrullinated fibrinogen β peptide and C3 deposition in rheumatoid arthritis patients. In addition, we characterized the humoral immune response against the viral EBNA-1 antigen to demonstrate the applicability of this assay beyond autoimmune conditions. We observed that particular buffer compositions were demanded for separate measurement of antibody reactivity and complement activation, as detection of antigen-antibody complexes appeared to be masked due to C3 deposition. We also found that rheumatoid factors of IgM isotype altered C3 deposition and introduced false-positive reactivities against EBNA-1 antigen. In conclusion, the presented bead-based assay setup can be utilized to profile antibody reactivities and immune-complex induced complement activation in a high-throughput manner and could facilitate the understanding and diagnosis of several diseases where complement activation plays role in the pathomechanism

Anti-citrullinated protein antibodies cause arthritis by cross-reactivity to joint cartilage

Today, it is known that autoimmune diseases start a long time before clinical symptoms appear. Anti-citrullinated protein antibodies (ACPAs) appear many years before the clinical onset of rheumatoid arthritis (RA). However, it is still unclear if and how ACPAs are arthritogenic. To better understand the molecular basis of pathogenicity of ACPAs, we investigated autoantibodies reactive against the C1 epitope of collagen type II (CII) and its citrullinated variants. We found that these antibodies are commonly occurring in RA. A mAb (ACC1) against citrullinated C1 was found to cross-react with several noncitrullinated epitopes on native CII, causing proteoglycan depletion of cartilage and severe arthritis in mice. Structural studies by X-ray crystallography showed that such recognition is governed by a shared structural motif "RG-TG" within all the epitopes, including electrostatic potential-controlled citrulline specificity. Overall, we have demonstrated a molecular mechanism that explains how ACPAs trigger arthritis

Evaluation of serum MMP-9 as predictive biomarker for antisense therapy in Duchenne

Duchenne Muscular Dystrophy (DMD) is a severe muscle disorder caused by lack of dystrophin. Predictive biomarkers able to anticipate response to the therapeutic treatments aiming at dystrophin re-expression are lacking. The objective of this study is to investigate Matrix Metalloproteinase-9 (MMP-9) as predictive biomarker for Duchenne. Two natural history cohorts were studied including 168 longitudinal samples belonging to 66 patients. We further studied 1536 samples obtained from 3 independent clinical trials with drisapersen, an antisense oligonucleotide targeting exon 51: an open label study including 12 patients; a phase 3 randomized, double blind, placebo controlled study involving 186 patients; an open label extension study performed after the phase 3. Analysis of natural history cohorts showed elevated MMP-9 levels in patients and a significant increase over time in longitudinal samples. MMP-9 decreased in parallel to clinical stabilization in the 12 patients involved in the open label study. The phase 3 study and subsequent extension study clarified that the decrease in MMP-9 levels was not predictive of treatment response. These data do not support the inclusion of serum MMP-9 as predictive biomarker for DMD patients

A retrospective analysis of cases with acute poisoning in Zonguldak, Turkey

Aim: The aim of this study was to evaluate etiological, demographical and clinical characteristics of cases with acute poisoning referred to the Emergency Medical Service (EMS) of Zonguldak Karaelmas University Hospital In Zonguldak, a City in the West Black Sea Region of Turkey. Materials and Methods: Two-hundred and ninety-five cases with acute intoxication referred to the Emergency Department of Zonguldak Karaelmas University Hospital between March 2003 and December 2006 were analyzed retrospectively. Result

Acute effects of pentobarbital, thiopental and urethane on lung oedema induced by alpha-naphthythiourea (ANTU)

This study was designed to investigate the possible participation of urethane, pentobarbital sodium and thiopental sodium anaesthesia in the lung oedema induced by alpha-naphthylthiourea (ANTU), which is a well known noxious chemical agent in the lung. ANTU when injected intraperitoneally (i.p.) into rats (10 mg kg-1 i.p.) produced lung oedema as indicated by an increase in lung weight/body weight (LW/BW) ratio and pleural effusion (PE) reaching a maximum within 4 h. Administration of urethane prior to ANTU, at doses of 100 and 200 mg (100 g)-1, elicited a significant and dose-dependent inhibition in LW/BW ratio and PE. Thiopental sodium at doses of 25, 50 mg kg-1, also produced a significant and dose-dependent inhibition of both parameters. Prior i.p. injection of pentobarbital sodium at a dose of 40 mg kg-1 elicited a significant inhibition in both parameters. These results suggest that i.p. urethane, thiopental sodium and pentobarbital sodium pretreatment have a prophylactic effect on ANTU-induced lung injury in rats. The possible role of the anaesthetics in lung oedema induced by ANTU and the possible underlying mechanisms are discussed. © 2002 Elsevier Science Ltd

Selenium, copper, zinc, iron levels and mortality in patients with sepsis and systemic inflammatory response syndrome in Western Black Sea Region, Turkey

Objective: To evaluate the changing levels of selenium, copper, zinc and iron in patients with sepsis and systemic inflammatory response syndrome and their influence on mortality. Methods: The prospective study was conducted at a tertiary care university hospital in Zonguldak city in the western Black Sea region of Turkey from January 2012 to December 2013, and comprised patients with sepsis and systemic inflammatory response syndrome. Blood samples were taken on 1st, 3rd, 5th and 7th days to measure serum selenium, copper, zinc and iron levels. Patients' demographic data, presence of additional diseases and mortality were recorded. Results: Of the 57 patients, 28(49.1%) were female and 29(50.9%) were male, with an overall mean age of 60.3±19.4 years, mean height of 166.1±11.4cm, mean weight of 76.5±17.5kg. Copper and zinc levels were in the normal range, while selenium and iron levels were lower than the limit values at all measuring periods. There was no significant difference between first and other days in accordance with element levels (p>0.05). Baseline copper levels in patients with malignancy were lower than patients without malignancy (p<0.05). In hypertensive patients, baseline copper levels were higher and 7th day levels were lower than non-hypertensive (p<0.05). Baseline selenium levels of those who died were lower than the other patients (p<0.05). Selenium and iron levels were decreased in patients with sepsis-systemic inflammatory response syndrome and copper levels were lower in patients with malignancy, hypertension and chronic obstructive pulmonary disease (p<0.05). There was no change in zinc levels of the patients. Conclusion: Reduced basal selenium levels of patients with sepsis and systemic inflammatory response syndrome were associated with mortality. © 2016, Pakistan Medical Association. All rights reserved

Effects of various anesthesia maintenance on serum levels of selenium, copper, zinc, iron and antioxidant capacity [Efeitos da manutenção de várias anestesias sobre os níveis séricos de selênio, cobre, zinco e ferro e a capacidade antioxidante]

Background and objectives: In this study, we aimed to investigate the effects of sevoflurane, desflurane and propofol maintenances on serum levels of selenium, copper, zinc, iron, malondialdehyde, and glutathion peroxidase measurements, and antioxidant capacity. Methods: 60 patients scheduled for unilateral lower extremity surgery which would be performed with tourniquet under general anesthesia were divided into three groups. Blood samples were collected to determine the baseline serum levels of selenium, copper, zinc, iron, malondialdehyde and glutathion peroxidase. Anesthesia was induced using 2-2.5 mg kg-1 propofol, 1 mg kg-1 lidocaine and 0.6 mg kg-1 rocuronium. In the maintenance of anesthesia, under carrier gas of 50:50% O2:N2O 4Lmin-1, 1 MAC sevoflorane was administered to Group S and 1 MAC desflurane to Group D; and under carrier gas of 50:50% O2:air 4 L min-1 6 mg kg h-1 propofol and 1µg kg h-1 fentanyl infusion were administered to Group P. At postoperative blood specimens were collected again. Results: It was observed that only in Group S and P, levels of MDA decreased at postoperative 48th hour; levels of glutathion peroxidase increased in comparison to the baseline values. Selenium levels decreased in Group S and Group P, zinc levels decreased in Group P, and iron levels decreased in all three groups, and copper levels did not change in any groups in the postoperative period. Conclusion: According to the markers of malondialdehyde and glutathion peroxidase, it was concluded that maintenance of general anesthesia using propofol and sevoflurane activated the antioxidant system against oxidative stress and using desflurane had no effects on oxidative stress and antioxidant system. © 2014 Sociedade Brasileira de Anestesiologia

Functional outcomes of multilevel botulinum toxin and comprehensive rehabilitation in cerebral palsy

The objective of this study was to measure the effect of lower extremity multilevel botulinum toxin A injections and comprehensive rehabilitation on spasticity and to determine the functional gains in ambulatory children with cerebral palsy. Sixteen ambulatory children with spastic cerebral palsy (9 hemiplegic, 7 diplegic), aged between 3 and 8 years, who were able to walk with or without assistance (Gross Motor Functional Classification System I-III) were recruited to the study. Botulinum toxin A injections were applied to a total of 23 extremities, followed by a comprehensive rehabilitation program. Walking distance and walking speed (evaluated by the Six-Minute Walk Test) were significantly improved after treatment. Similarly, scores on the Observational Gait Scale (assessed by video gait analysis) increased significantly. Improvements in muscle length, spasticity, and selectivity were recorded. Reduced muscle spasticity after botulinum toxin A injections in children with cerebral palsy, with a comprehensive rehabilitation program, enabled clinically relevant improvements in functional ability. © 2011 The Author(s)

Antimicrobial effects of two anaesthetic agents: Dexmedetomidine and midazolam

Some anaesthetic agents are known to inhibit microbial growth. The aim of this in vitro study was to investigate possible antimicrobial effects of two frequently used agents in intensive care units, dexmedetomidine and midazolam. Antimicrobial effect was tested on Staphylococcus aureus, Enterococcus faecalis, Escherichia coli and Pseudomonas aeruginosa by broth microdilution method. Midazolam showed inhibitor and bactericidal effect on S. aureus at concentrations 256 µg.ml-1 and 512 µg.ml-1 respectively and on E. faecalis at concentrations 128 µg.ml-1 and 256 µg.ml-1. Dexmedetomidine demonstrated inhibitor effect on S. aureus, E. coli and P. aeruginosa at concentrations 32 µg.ml-1, 16 µg.ml-1 and 16 µg.ml-1 respectively. Midazolam had inhibitor and bactericidal effects on S. aureus and E. faecalis. Dexmedetomidine had only inhibitor effects on S. aureus, E. coli and P. aeruginosa. Further studies are needed to determine the antimicrobial mechanisms and clinical applications

Factors associated with postoperative mortality in geriatric orthopedic surgery: A retrospective analysis of single center data

Objective: In our study the factors related to anesthesia and peroperative variables associated with postoperative mortality among patients aged ?65 years who had undergone orthopedic surgery were assessed. Methods: Reports on patients aged ?65 years who had undergone orthopedic surgery between 2015 and 2017 were investigated retrospectively. Results: A total of 135 patients were included in the study. The operations comprised implantations of total hip prosthesis in 26%, total knee prosthesis in 18%, fixation of lower extremity fractures in 24, and upper extremity fractures in 14%, and amputation surgery in 17% of the patients. The postoperative mortality rates were highest (76.9%) among patients who underwent amputation surgery (p<0.05). It was found that anesthesia type, whether regional or general, was not related to mortality. Mortality was found to be associated with increasing age, ?3 ASA score, emergency surgery, ?3 accompanying diseases, prolonged preoperative hospital stay and low preoperative hemoglobin (Hb) values (p<0.05). Patients developing postoperative complications, those who were monitored in intensive care unit (ICU) and required mechanical ventilator (MV), and patients with prolonged ICU and hospital stay had higher mortality rates (p<0.05). 9% of all patients were determined dead. Conclusion: Among geriatric orthopedic surgery patients, apart from gender and anesthesia method, increasing age, high ASA scores, emergency surgery, the number of accompanying diseases, duration of preoperative hospital stays, low preoperative Hb values, postoperative complications requiring ICU-MV and prolonged ICU and hospital stays were all factors that affected postoperative mortality. We believe that detailed preoperative assessment and perioperative clinical management are essential if postoperative prognosis after geriatric orthopedic surgery is to be improved. © 2019 Copyright Anesthesiology and Reanimation Specialists' Society