Experience of having a sibling with autism spectrum disorder: a systematic review protocol.

OBJECTIVE: This qualitative systematic review aims to understand the experiences of neurotypical siblings of a person with autism spectrum disorder. INTRODUCTION: Autism spectrum disorder influences communication and social interaction with other people and has a significant impact on family relationships. The experiences of siblings range from the positive, such as development of increased empathy and ability to cope with challenges, to experiences that are more negative, such as a higher risk of bullying. In many countries, neurotypical siblings are marginalized and don't receive adequate support to cope with the increased challenges. INCLUSION CRITERIA: This review will consider qualitative studies exploring the experiences of siblings of a person with autism spectrum disorder. There will be no limitations regarding age, gender, sex, or length of relationship with the sibling. We will consider studies from all countries and contexts. METHODS: This study will be conducted according to the JBI methodology for qualitative reviews. A 3-step search strategy will be used to find published and unpublished studies from the following sources: MEDLINE, CINAHL, APA PsycINFO, Scopus, SocINDEX, Web of Science, Embase, ERIC, ProQuest Dissertations and Theses, Open Dissertations, and Google Scholar (first 100 records). There will be no search limitations on the publication period or language, but only studies with an English-language abstract/title will be considered for inclusion. Screening, data extraction, and data synthesis will be conducted by 2 independent reviewers.

Impact of educational and psychological interventions compared with standard care in college/university students with autism spectrum disorder: a systematic review protocol.

OBJECTIVE: This review will explore the impact of educational and psychological interventions on educational, social, behavioral, and mental health outcomes in students with autism spectrum disorder in tertiary education. INTRODUCTION: This systematic review will inform a new guideline on support for students with autism spectrum disorder in the tertiary education environment. These students face multiple educational, behavioral, social, and health-related problems that require effective interventions. INCLUSION CRITERIA: Participants are students with autism spectrum disorder in a tertiary education study program. Educational and psychological interventions will be included, such as accommodations, meta-cognitive and self-regulation training, psychological counseling, social skills training, and peer-mentoring/academic coaching. The comparator will be standard care. The study outcomes will include academic drop-out rates and evaluations, learning skills, social skills and social engagement, behavior, mental health (including anxiety, stress, and depression), and employment after graduation. This review will consider quantitative studies only. METHODS: A 3-step search strategy will be used to find both published and unpublished studies in MEDLINE, CINAHL, APA PsycINFO, SocINDEX, Web of Science, Clinical Trials, ProQuest Dissertations and Theses, Open Dissertations, ERIC, WHO ICRTP, and Google Scholar. There will be no date or language limitations. All stages of article screening, critical appraisal, and data extraction will be conducted by 2 independent reviewers with the resolution of any disagreements done via consensus or through a senior reviewer. If possible, the included studies will be pooled using meta-analysis. The degree of certainty of the evidence will be assessed according to the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) approach. SYSTEMATIC REVIEW REGISTRATION NUMBER: PROSPERO CRD42022323554.

Identification of telehealth nursing approaches in the light of the COVID-19 pandemic-A literature review.

AIMS: This study aimed to identify recommendations for quality nursing care provision, focusing on the possibilities of delivering telehealth nursing at the national level in the Czech Republic. BACKGROUND: The significant growth in the use of technology in health care has changed the environment for patient care and how health care is provided. The COVID-19 pandemic has shown the requirement for telemedicine use in everyday clinical practice. EVALUATION: A literature review aims to find guidelines, recommendations, manuals, standards or consensus papers published in 2017-2022. KEY ISSUES: In total, 12 guidelines were identified. Based on a critical evaluation carried out by two experts, the World Health Organization guideline focused on digital interventions to strengthen the health system has been identified. This guideline was evaluated to be of the highest quality and in line with the Grading of Recommendations, Assessment, Development and Evaluations methodology. CONCLUSION: The provision of telemedicine nursing interventions is currently limited at the national level and is not entirely accepted in reimbursement yet. The interprofessional collaboration will be initiated to make recommendations for specific patient groups. IMPLICATIONS FOR NURSING MANAGEMENT: Selected guidelines can be recommended for national adoption to set up and support nursing interventions in telemedicine, including nursing management settings.

Corrigendum: Tactile low frequency vibration in dementia management: A scoping review.

[This corrects the article DOI: 10.3389/fpsyg.2022.854794.].

Monoclonal anti-endoglin antibody TRC105 (carotuximab) prevents hypercholesterolemia and hyperglycemia-induced endothelial dysfunction in human aortic endothelial cells.

Endoglin (Eng) is a co-receptor of the transforming growth factor ß superfamily playing an important role in endothelial dysfunction. TRC105 (carotuximab) is a monoclonal antibody that blocks Eng and its downstream Smad signaling pathway. Here we have investigated for the first time the effects of TRC105 treatment on the development of endothelial dysfunction induced by 7-ketocholesterol (7K) or high glucose (HG), focusing on Eng expression, signaling, and function. In the hypercholesterolemia study, human aortic endothelial cells (HAoECs) were treated with TRC105 (300 µg/ml) for 1 h, followed by the addition of 7K (10 µg/ml) for another 12 h. In the hyperglycemia study, HAoECs were exposed to HG (45 mM) for 60 h, followed by the addition of TRC105 for another 12 h, and cells treated with 5mM glucose and 40 mM mannitol served as control. Protein levels, adhesion, and transmigration of monocytes were assessed by flow cytometry, mRNA expression was measured by qRT-PCR. 7K and HG treatment increased protein levels of NF-κB and Eng and adhesion and transmigration of monocytes through HAoECs monolayer. TRC105 pretreatment reduced the 7K- or HG-induced Eng protein levels and pSmad1/5 and pSmad2/3 signaling. Despite increased protein levels of P-selectin and VCAM-1, TRC105 mediated blockage of Eng prevented 7K- and HG-induced adhesion and transmigration of monocytes through endothelial monolayers. These results suggest that TRC105-mediated Eng blockage can counteract the hypercholesterolemia- and hyperglycemia-induced endothelial dysfunction in HAoECs, suggesting that Eng might be a potential therapeutic target in disorders associated with elevated cholesterol and glucose levels.

Tactile Low Frequency Vibration in Dementia Management: A Scoping Review.

The prevalence of dementia is increasing with the ever-growing population of older adults. Non-pharmacological, music-based interventions, including sensory stimulation, were reported by the Lancet Commission in 2020 to be the first-choice approach for managing the behavioural and psychological symptoms of dementia. Low frequency sinusoidal vibration interventions, related to music interventions through their core characteristics, may offer relief for these symptoms. Despite increasing attention on the effectiveness of auditory music interventions and music therapy for managing dementia, this has not included low frequency vibration. This scoping review, following the JBI methodology guidelines, was conducted to investigate participants' responses to both sound and mechanical vibration, the characteristics of the delivered interventions, methodological challenges, and the specifics of the research experiments reported. An extensive search was conducted in BMC, CINAHL, Cochrane Central Register of Controlled Trials, EMBASE, ERIC, MEDLINE (OvidSP), Pedro, ProQuest Central, PsycINFO, Scopus, and Web of Science. Current Controlled Trials, Clinical Trials, and Google Scholar were also searched as well as a hand search in relevant journals. Studies on adults with all types of dementia, investigating tactile low frequency sound or mechanical vibration in any context were considered. Data from eight full-length studies (three RCTs, two quasi-experimental, two case reports, and one qualitative) were extracted using the data extraction table developed by the authors and were included in the analysis and critical appraisal. Issues in quality related to, for example, control groups and blinding. Few studies addressed participants' subjective responses to the interventions. Reporting on the intervention characteristics was unclear. It appeared more frequent sessions led to better outcomes and home-based interventions potentially addressing the issue of access and feasibility. Future research should include neuroimaging to measure and confirm the hypothesised mechanism of cerebral coherence. Standardised reporting of intervention characteristics is also needed to ensure replicability of the experiments. Higher quality research is needed to investigate the impact and effect of low frequency vibration for the symptoms of dementia and compare outcomes in meta-syntheses.

Metabolomics of Cerebrospinal Fluid in Multiple Sclerosis Compared With Healthy Controls: A Pilot Study.

Background: Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system (CNS) leading to the loss of myelin and axons. Diagnosis is based on clinical findings, MRI, and analysis of cerebrospinal fluid (CSF). CSF is an ultrafiltrate of plasma and reflects inflammatory processes in the CNS. The aim of this study was to perform metabolomics analysis of CSF in patients after the first attack of MS and healthy controls and try to find new specific analytes for MS including those potentially predicting disease activities at the onset. Methods: We collected CSF from 19 patients (16 females, aged 19-55 years) after the first attack of clinical symptoms who fulfilled revised McDonald criteria of MS and CSF of 19 controls (16 females, aged 19-50 years). Analyses of CSF samples were provided using the high-performance liquid chromatography system coupled with a mass spectrometer with a high-resolution detector (TripleTOF 5600, AB Sciex, Canada). Results: Approximately 130 selected analytes were identified, and 30 of them were verified. During the targeted analysis, a significant decrease in arginine and histidine and a less significant decrease in the levels of asparagine, leucine/isoleucine, and tryptophan, together with a significant increase of palmitic acid in the patient group, were found. Conclusion: We observed significant differences in amino and fatty acids in the CSF of newly diagnosed patients with MS in comparison with controls. The most significant changes were observed in levels of arginine, histidine, and palmitic acid that may predict inflammatory disease activity. Further studies are necessary to support these findings as potential biomarkers of MS.

The Effects of Ayres Sensory Integration and Related Sensory Based Interventions in Children with Cerebral Palsy: A Scoping Review.

The theory of Ayres Sensory Integration® was formulated in the 1960s, and is also known as sensory integration (SI). It has been used in people with cerebral palsy (CP), though the research evidence for its effects in this population is contradictory and inconclusive. To fill in this knowledge gap, we conducted a scoping review of the body of literature on the topic, including any type of quantitative or qualitative research of SI in people with CP without any restrictions of age, language, geography, professionals involved, etc. In September 2020, we searched Scopus, ProQuest Central, MEDLINE (via PubMed), CINAHL Plus and the Academic Search Ultimate and Web of Science, as well as the grey literature sources OpenGrey and MedNar. Two reviewers independently screened the texts and the references lists of the included papers. We finally included seven relevant papers (four randomized controlled trials, two quasi-experimental studies and one case series), though not all fidelity measures required for Ayres SI were reported in the papers. The age of participants ranged from 3 months to 15 years; no studies were identified on adults. There is some evidence that SI or related sensory-based interventions (SBI) may be useful for movement development and other outcomes (attention span, therapy of sensory processing disorders, body perception and therapy of strabismus), but there is only scarce and low-quality evidence comparing interventions. We recommend to conduct well-designed randomized controlled trials (RCTs) with an optimal sample size on the effectiveness of formal Ayres SI for the motor development or other outcomes (as attention span or self-care abilities) using standardized measurement tools.

Affiblot: a dot blot-based screening device for selection of reliable antibodies.

The key factor in the development of antibody-based assays is to find an antibody that has an appropriate affinity, high specificity, and low cross-reactivity. However, this task is not easy to carry out since the research antibodies on the market may suffer from low specificity and reproducibility. Here, we report on a palm-sized dot blot-based device, called the affiblot, that has a specially designed lid that allows simultaneous semi-quantitative comparison of up to five antibodies from different suppliers regarding their affinity/avidity, cross-reactivity, and batch-to-batch reliability. The only required peripheral equipment is a vacuum pump, a camera, and densitometry software. The affiblot device was tested for its functionality and its measurements were compared against those obtained by standard dot blot and ELISA. The benefit over these methods, when various antibodies are evaluated, is in its simplicity. It allows easy antigen deposition, fast application and the discarding of the solutions, a compact undivided membrane, and therefore significant decrease of labor. The device was tested with specific anti-ApoE, anti-EpCAM, anti-Salmonella, anti-E. coli, and anti-Listeria antibodies from different suppliers. Their properties were compared for their ability to interact specifically with antigen and/or non-target structures and the best-suited antibody for the intended application was identified.

Adaptation of Music Therapists' Practice to the Outset of the COVID-19 Pandemic-Going Virtual: A Scoping Review.

Background: In the midst of a worldwide COVID-19 pandemic, music therapists previously not involved in telehealth had to develop effective remote forms of music therapy. The objective of this review was to systematically explore how music therapists previously working in-person adapted to the transfer to remote forms of therapy in the context of the coronavirus outbreak. Methods: We searched Scopus, Web of Science Core Collection, CINAHL, Medline, ProQuest Central, PubMed, EMBASE, PsycINFO and PsyARTICLES, grey literature (to October 2020), and websites of professional organizations. We followed the JBI methodology for scoping reviews. Results: Out of the 194 screened texts, we included ten very heterogeneous articles with an overall very low quality. Most texts described remote therapy in the form of synchronous video calls using the Internet, one paper described a concert in a patio of a residential home. We report the authors' experience with the adaptation and activities, challenges and benefits of remote forms of therapy, recommendations of organizations, and examples and tips for online therapies. Conclusions: Music therapists have adapted the musical instruments, the hours, the technology used, the therapeutic goals, the way they prepared their clients for sessions, and other aspects. They needed to be more flexible, consult with colleagues more often, and mind the client-therapist relationship's boundaries. It seems, when taken as a necessary short-term measure, online music therapy works sufficiently well. The majority of papers stated that benefits outweighed the challenges, although many benefits were directly linked with the pandemic context.

Tactile Low Frequency Vibration in Dementia Management: A Scoping Review Protocol.

Dementia is a growing issue in modern society. Non-pharmacological interventions such as music are suggested as the primary methods for symptom management. Therapeutic potential may also be found in sound/mechanical low frequency vibrations (LFV) that share the core characteristics of music, but these are lesser understood. The aim of the proposed scoping review is to explore the responses of persons with dementia to LFV, e.g., vibroacoustic therapy or whole-body vibration. The scoping review will follow the Joanna Briggs Institute methodology guidelines. An extensive search in BMC, CINAHL, Cochrane Central Register of Controlled Trials, EMBASE, ERIC, MEDLINE (OvidSP), Pedro, ProQuest Central, PsycINFO, Scopus, Web of Science, and grey literature sources in Clinical Trials, Current Controlled Trials, Google Scholar, and manual search of relevant journals is planned to find all relevant research papers. The paper selection, full-text assessment, and data extraction will be performed by two independent reviewers. Participants' responses to the interventions and the experiment designs, including methodological challenges, will be analysed and compared. Results may highlight potential gaps in reporting and comparing sound and mechanical vibration approaches and promote better understanding of their potential for managing the symptoms of dementia. Furthermore, the possible relationships between LFV and music-based interventions may become clearer.

Arts Therapies Interventions and Their Outcomes in the Treatment of Eating Disorders: Scoping Review Protocol.

Arts therapies (AsTs) are considered a valuable intervention for people with eating disorders, however the range of research studies and the comparison between the types of arts therapies are unknown. The goal of the future scoping review is to explore the therapeutic outcomes addressed by arts therapists in research studies on people with eating disorders and compare the different types of arts-based interventions. This scoping review will be conducted in accordance with the Joanna Briggs Institute methodology. Included will be research studies and sources oriented towards people with eating disorders of all ages and AsTs of any type (art therapy, drama therapy, music therapy, dance/movement therapy, and expressive therapies). There is no language/publication period limitation. The following databases will be searched: CINAHL Plus, EMBASE, MEDLINE (OvidSP), ProQuest Central, PsycINFO, PubMed, Scopus, and Web of Science. Sources of unpublished studies and grey literature will include Google Scholar, MedNar, clinical trials, and current controlled trials. Titles/abstracts and full texts of studies will be assessed against the inclusion criteria, and the data extracted by two independent reviewers. Based on the results, we will compare the types of AsTs according to the research designs, country/settings, intervention methods/materials, adverse effects reported, and therapeutic outcomes related to AsTs.

Severe chronic lithium intoxication in patient treated for bipolar disorder.

OBJECTIVE: Lithium has been long used in psychiatry as an adjuvant treatment for bipolar disorder. Chronic lithium intoxication is very rare. DESIGN: We present the case of a 72-year-old female, treated with lithium for more than 10 years for bipolar disorder, who was admitted for gait impairment with weakness of limbs, myoclonus, speech impairment and memory disturbances. RESULTS: Diagnosis of lithium intoxication was based on clinical picture and determination of serum lithium levels. EEG showed severe encephalopathy with triphasic wave complexes. Sensory and motor axonal neuropathy was observed by EMG. Discontinuation of the drug leads to clinical improvement, although not to a fully neurological recovery. CONCLUSION: Lithium is still very effective drug, but requires regular monitoring of serum levels to prevent overdose and symptoms of intoxication. Neurophysiological methods, including EEG and EMG, are strongly recommended to determine the level of peripheral and/or central nervous system impairment.

Extrapontine myelinolysis manifested selectively by acute severe parkinsonian syndrome. Case Report.

OBJECTIVES: Osmotic demyelination syndrome (ODMS) is a rare and serious neurologic disorder with acute myelin disintegration, usually in the pontine area (central pontine myelinolysis) and to a lesser extent, even in other areas of the central nervous system (extrapontine myelinolysis). The main underlying mechanism is the change of serum osmolality with quick correction of low mineral levels, mainly hyponatraemia. Clinical manifestation is various and depends on the localization. DESIGN: We describe an acute isolated extrapontine myelinolysis causing acute onset of parkinsonism in a 61-year-old man who developed quickly progressing parkinsonian syndrome after the rapid correction of hyponatraemia. RESULTS: Brain MRI revealed lesions only in the striatum, sparing the globus pallidus. Substitution therapy with high doses of levodopa significantly improved his clinical condition. CONCLUSION: Extrapontine myelinolysis with isolated affection of basal ganglia is extremely rare. In such case, clinical manifestation of acute severe parkinsonism could be successfully treated by high dose of levodopa.

An integrated microfluidic chip for immunocapture, preconcentration and separation of ß-amyloid peptides.

We present an integrated microfluidic chip for detection of ß-amyloid (Aß) peptides. Aß peptides are major biomarkers for the diagnosis of Alzheimer's disease (AD) in its early stages. This microfluidic device consists of three main parts: (1) An immunocapture microcolumn based on self-assembled magnetic beads coated with antibodies specific to Aß peptides, (2) a nano-porous membrane made of photopolymerized hydrogel for preconcentration, and (3) a microchip electrophoresis (MCE) channel with fluorescent detection. Sub-milliliter sample volume is either mixed off-chip with antibody coated magnetic beads and injected into the device or is injected into an already self-assembled column of magnetic beads in the microchannel. The captured peptides on the beads are then electrokinetically eluted and re-concentrated onto the nano-membrane in a few nano-liters. By integrating the nano-membrane, total assay time was reduced and also off-chip re-concentration or buffer exchange steps were not needed. Finally, the concentrated peptides in the chip are separated by electrophoresis in a polymer-based matrix. The device was applied to the capture and MCE analysis of differently truncated peptides Aß (1-37, 1-39, 1-40, and 1-42) and was able to detect as low as 25 ng of synthetic Aß peptides spiked in undiluted cerebrospinal fluid (CSF). The device was also tested with CSF samples from healthy donors. CSF samples were fluorescently labelled and pre-mixed with the magnetic beads and injected into the device. The results indicated that Aß1-40, an important biomarker for distinguishing patients with frontotemporal lobe dementia from controls and AD patients, was detectable. Although the sensitivity of this device is not yet enough to detect all Aß subtypes in CSF, this is the first report on an integrated or semi-integrated device for capturing and analyzing of differently truncated Aß peptides. The method is less demanding and faster than the conventional Western blotting method currently used for research.

PEGylation of magnetic poly(glycidyl methacrylate) microparticles for microfluidic bioassays.

In this study, magnetic poly(glycidyl methacrylate) microparticles containing carboxyl groups (PGMA-COOH) were coated using highly hydrophilic polymer poly(ethylene glycol) (PEG). PEG was used to reduce nonspecific interactions with proteins and cells while decreasing adhesion of particles to the walls of a microfluidic devices from poly(dimethylsiloxane) (PDMS) and cyclic olefin copolymer (COC). Zeta potential measurement, infrared spectroscopy, scanning electron microscopy, anti-PEG ELISA assay, and bioaffinity interactions between biotin and streptavidin-HRP successfully proved the presence of PEG on the surface of microspheres. Both neat and PEGylated microspheres were then incubated with the inert protein bovine serum albumin or cells to evaluate the rate of nonspecific adsorption (NSA). PEG with Mr of 30,000 Da was responsible for 45% reduction in NSA of proteins and 74% for cells compared to neat particles. The microspheres' behavior in PDMS and COC microchannels was then evaluated. Aggregation and adhesion of PEGylated microspheres significantly decreased compared to neat particles. Finally, the model enzyme horseradish peroxidase was immobilized on the microspheres through the heterobifunctional PEG chain. The possibility for subsequent covalent coupling of the ligand of interest was confirmed. Such PEGylated microparticles can be efficiently used in PDMS microchips as a carrier for bioaffinity separation or of enzyme for catalysis.

Application of an improved magnetic immunosorbent in an Ephesia chip designed for circulating tumor cell capture.

In this study, we describe a particular step in developing a microfluidic device for capture and detection of circulating tumor cells-specifically the preparation of an immunosorbent for implementation into the separation chip. We highlight some of the most important specifics connected with superparamegnetic microspheres for microfluidic purposes. Factors such as nonspecific adsorption on microfluidic channels, interactions with model cell lines, and tendency to aggregation were investigated. Poly(glycidyl methacrylate) microspheres with carboxyl groups were employed for this purpose. To address the aforementioned challenges, the microspheres were coated with hydrazide-PEG-hydrazide, and subsequently anti-epithelial cell adhesion molecule (EpCAM) antibody was immobilized. The prepared anti-EpCAM immunosorbent was pretested using model cell lines with differing EpCAM density (MCF7, SKBR3, A549, and Raji) in a batchwise arrangement. Finally, the entire system was implemented and studied in an Ephesia chip and an evaluation was performed by the MCF7 cell line.

Albumin-coated monodisperse magnetic poly(glycidyl methacrylate) microspheres with immobilized antibodies: application to the capture of epithelial cancer cells.

Monodisperse (4 µm) macroporous crosslinked poly(glycidyl methacrylate) (PGMA) microspheres for use in microfluidic immunomagnetic cell sorting, with a specific application to the capture of circulating tumor cells (CTCs), were prepared by multistep swelling polymerization in the presence of cyclohexyl acetate porogen and hydrolyzed and ammonolyzed. Iron oxide was then precipitated in the microspheres to render them magnetic. Repeated precipitation made possible to raise the iron oxide content to more than 30 wt %. To minimize nonspecific adsorption of the microspheres in a microchannel and of cells on the microspheres, they were coated with albumin crosslinked with glutaraldehyde. Antibodies of epithelial cell adhesion molecule (anti-EpCAM) were then immobilized on the albumin-coated magnetic microspheres using the carbodiimide method. Capture of breast cancer MCF7 cells as a model of CTCs by the microspheres with immobilized anti-EpCAM IgG was performed in a batch experiment. Finally, MCF7 cells were captured by the anti-EpCAM-immobilized albumin-coated magnetic microspheres in an Ephesia chip. A very good rejection of lymphocytes was achieved. Thus, albumin-coated monodisperse magnetic PGMA microspheres with immobilized anti-EpCAM seem to be promising for capture of CTCs in a microfluidic device.

Development of a magnetic immunosorbent for on-chip preconcentration of amyloid ß isoforms: Representatives of Alzheimer's disease biomarkers.

Determination of amyloid ß (Aß) isoforms and in particular the proportion of the Aß 1-42 isoform in cerebrospinal fluid (CSF) of patients suspected of Alzheimer's disease might help in early diagnosis and treatment of that illness. Due to the low concentration of Aß peptides in biological fluids, a preconcentration step prior to the detection step is often necessary. This study utilized on-chip immunoprecipitation, known as micro-immunoprecipitation (µIP). The technique uses an immunosorbent (IS) consisting of magnetic beads coated with specific anti-Aß antibodies organized into an affinity microcolumn by a magnetic field. Our goal was to thoroughly describe the critical steps in developing the IS, such as selecting the proper beads and anti-Aß antibodies, as well as optimizing the immobilization technique and µIP protocol. The latter includes selecting optimal elution conditions. Furthermore, we demonstrate the efficiency of anti-Aß IS for µIP and specific capture of 5 Aß peptides under optimized conditions using various subsequent analytical methods, including matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), capillary electrophoresis, microchip electrophoresis, and immunoblotting. Synthetic Aß peptides samples prepared in buffer and spiked in human CSF were analyzed. Finally, on-chip immunoprecipitation of Aß peptides in human CSF sample was performed.

Identification of carbonic anhydrase I immunodominant epitopes recognized by specific autoantibodies which indicate an improved prognosis in patients with malignancy after autologous stem cell transplantation.

This work employs an epitope mapping of carbonic anhydrase (CA), isoform I (CA I), for detection of the main immunodominant epitopes. Our interest has arisen from an observed spontaneous tumor regression in patients who developed an aplastic anemia type syndrome after a high-dose therapy with autologous stem cell transplantation and whose sera contained high titer of anti carbonic anhydrase (anti-CA) autoantibodies. There are many indications that the presence of these autoantibodies may provide significant survival benefit for the patients. Western blot analysis confirmed strong immunoreactivity of the patients' sera with several CA isoforms and the CA I has been selected for our study as a highly abundant and widely distributed isoform. The applied analytical approach consists of specific fragmentation of CA I protein followed by immunospecific isolation of peptides reacting with polyclonal anti-CA I autoantibodies of patients in spontaneous remission. We improved the standard epitope mapping schema by incorporating the benefits of magnetic carriers and biomagnetic separation techniques. Mass spectrometry has been applied for detection and identification of epitopes and the acquired results were verified by bioinformatic tools. The candidate epitopes of CA I (NVGHS, DGLAV, SSEQL, and SLKPI) are discussed herein as potential therapeutic targets. This work highlights the usefulness of the epitope mapping technique based on magnetic microspheres for effective and rapid determination of immunodominant epitopes of the target protein.