Složen protokol liječenja bolesnika s amiotrofičnom lateralnom sklerozom

Amyotrophic lateral sclerosis is a progressive and fatal degenerative neuromuscular disease with few if any treatment options and physical rehabilitation addressing specific deficits is the most frequent form of therapy. Patients also suffer from depression and increased anxiety. Our purpose was to assess the neurorehabilitation effectiveness in a patient with amyotrophic lateral sclerosis who underwent stem cell transplantation but refused physiotherapy due to depression. Disease progression was followed using the revised Amyotrophic Lateral Sclerosis Functional Rating Scale bimonthly for six months pre- and then post-stem cell transplantation. Psychological traits were assessed using six standardized tests. Quantitative electroencephalogram diagnostics was performed before the first and after the last neurofeedback session, and sessions were conducted on a 3-times-a-week basis. The physiotherapy protocol included proprioceptive neuromuscular facilitation, electrical modalities unit applied to the lumbar spine area, and breathing, relaxation and walking exercises, among others. Increased motivation and marked decrease in the pain level was associated with the patient’s willingness to complete physiotherapy, which resulted in improvements in most neuromuscular deficits and in increased respiratory capacity. During the 12 post-rehabilitation months, progression of the disease decelerated, and a positive behavioral change was noted. The study suggested that neurofeedback could be used as a neurorehabilitation component of the personalized complex rehabilitation protocol in patients with amyotrophic lateral sclerosis.Amiotrofična lateralna skleroza je progresivna i smrtonosna degenerativna neuromuskularna bolest za koju postoji malo, ako uopće ijedna mogućnost liječenja pa je najčešći oblik terapije fizikalna rehabilitacija usmjerena na točno određene nedostatke. Ovi bolesnici pate i od depresije te pojačane anksioznosti. Cilj istraživanja bio je procijeniti učinkovitost neurorehabilitacije u bolesnika s amiotrofičnom lateralnom sklerozom koji je podvrgnut transplantaciji matičnih stanica, ali je zbog depresije odbio fizikalnu terapiju. Progresija bolesti praćena je pomoću revidirane Ljestvice za funkcionalnu ocjenu amiotrofične lateralne skleroze svaka dva mjeseca kroz šest mjeseci prije te nakon transplantacije matičnih stanica. Psihološke značajke procjenjivane su pomoću šest standardiziranih testova. Kvantitativna elektroencefalografska dijagnostika provedena je prije prvog i nakon posljednjeg neurofeedback tretmana, koji su se provodili tri puta na tjedan. Protokol fizikalne terapije obuhvaćao je, među ostalim, proprioceptivnu neuromuskularnu facilitaciju, jedinicu električnih modaliteta primijenjenu u području lumbalne kralježnice te vježbe disanja, opuštanja i hodanja. Pojačana motivacija i znatno sniženje razine boli bili su udruženi s bolesnikovim pristankom na potpunu fizikalnu terapiju, što je rezultiralo poboljšanjem većine neuromuskularnih nedostataka i povećanim dišnim kapacitetom. Tijekom 12 mjeseci nakon rehabilitacije progresija bolesti se usporila i zabilježena je pozitivna promjena ponašanja. Ovo istraživanje je pokazalo da se neurofeedback može primijeniti kao neurorehabilitacijska sastavnica personaliziranog složenog protokola rehabilitacije u bolesnika s amiotrofičnom lateralnom sklerozom

Comparison of printed glycan array, suspension array and ELISA in the detection of human anti-glycan antibodies

Anti-glycan antibodies represent a vast and yet insufficiently investigated subpopulation of naturally occurring and adaptive antibodies in humans. Recently, a variety of glycan-based microarrays emerged, allowing high-throughput profiling of a large repertoire of antibodies. As there are no direct approaches for comparison and evaluation of multi-glycan assays we compared three glycan-based immunoassays, namely printed glycan array (PGA), fluorescent microsphere-based suspension array (SA) and ELISA for their efficacy and selectivity in profiling anti-glycan antibodies in a cohort of 48 patients with and without ovarian cancer. The ABO blood group glycan antigens were selected as well recognized ligands for sensitivity and specificity assessments. As another ligand we selected P1, a member of the P blood group system recently identified by PGA as a potential ovarian cancer biomarker. All three glyco-immunoassays reflected the known ABO blood groups with high performance. In contrast, anti-P1 antibody binding profiles displayed much lower concordance. Whilst anti-P1 antibody levels between benign controls and ovarian cancer patients were significantly discriminated using PGA (p = 0.004), we got only similar results using SA (p = 0.03) but not for ELISA. Our findings demonstrate that whilst assays were largely positively correlated, each presents unique characteristic features and should be validated by an independent patient cohort rather than another array technique. The variety between methods presumably reflects the differences in glycan presentation and the antigen/antibody ratio, assay conditions and detection technique. This indicates that the glycan-antibody interaction of interest has to guide the assay selection

Optimal Combination of Glycan-Based Serum Diagnostic Markers Which Maximize AUC

Recently a new high-throughput biomarker discovery platform based on printed glycan arrays (PGA) has emerged. PGAs are similar to DNA arrays but contain deposits of various carbohy-drate structures (glycans) instead of spotted DNAs. PGA-based biomarker discovery for the early detection, diagnosis and prognosis of human malignancies is based on the response of the immune system as measured by the level of binding of anti-glycan antibodies from human serum to the glycans on the ar-ray. Since the PGA offer a multitude of markers which can have moderate individual diagnostic power they can be combined in order to achieve maximal classification precision assessed by the popular performance measure area under the ROC curve (AUC). This paper presents an empirical analysis of several combination approaches including those that are specifically designed to maximize the AUC and those that are not, such as Fisher Linear Discriminant, Support Vector Machines and Gen-eralized Linear Model. The analysis is performed on real-life PGA data from three pilot studies involving malignant mesothe-lioma, lung cancer and ovarian cancer

Serum antiglycan antibody detection of nonmucinous ovarian cancers by using a printed glycan array

Epithelial ovarian cancer has the highest mortality rate among gynecological cancers. Altered glycosylation is associated with oncogenic transformation producing tumor-associated carbohydrate antigens. We investigated the potential of natural occurring antiglycan antibodies in the diagnosis of ovarian cancer by using printed glycan array. Antiglycan antibodies bound to 203 chemically synthesized printed glycans were detected via biotin-streptavidin fluorescence system in serum of women with normal operative findings (healthy controls; n = 24) and nonmucinous borderline or ovarian cancer of various FIGO stages (n = 33). Data were validated measuring blood group associated di-, tri and tetrasaccharide antigens on known ABO blood groups. Antiglycan antibodies demonstrated high reproducibility (r(c) > 0.9). Cluster analysis identified repetitive patterns of specific core carbohydrate structures: 11 N-linked glycans, 3 O-linked glycans and 2 glycosphingolipids. Biomarker detection revealed 24 glycans including P-1 (Gal alpha 1-4Gal beta 1-4GlcNAc beta; p < 0.001) significantly discriminating between (low-) malignant tumors and healthy controls. Comparable sensitivity and specificity with tumor marker CA125 was achieved by a panel of multivariate selected and linear combined antiglycan antibody signals (79.2 and 84.8%, respectively). Our findings demonstrate the potential of glycan arrays in the development of a new generation of biomarkers for ovarian cancer

Natural anti-glycan IgM recognize P1 glycosphingolipid expressed on ovarian cancer cells

Recent research strongly suggests a role of plasma-derived anti-P₁ antibodies (AGA) in ovarian cancer, as demonstrated by three independent glycan-based immunoassays. The level of these antibodies was lower in ovarian cancer patients and therefore discriminate cancer patients from healthy women. Here we investigate in a separate Australian cohort (n=155) whether IgM or IgG to P₁ accounts for this discrimination and whether plasma matched ascites samples contain AGA. We also aimed to identify the P₁ antigen in cancer tissue and cultured cells that are recognized by naturally occurring anti-P₁ IgM and to investigate the potential function in respect to cell migration. Our results demonstrated that the IgM to P₁ discriminates patients with ovarian cancer from healthy controls (p=0.0002) and that lower anti-P₁ antibody levels are associated with a slightly higher risk for early relapse. Mass spectrometry identified P₁ and structurally related epitopes in fresh tissue specimens and cultured cells. Ovarian cancer cell line IGROV1 was identified to be P₁-positive while others (n=7) including human ovarian surface epithelial cells were negative determined by comprehensive flow cytometry. Epitope-mapped and characterized affinity purified anti-P₁ antibodies from ascites were shown to bind P₁-expressing cancer cells. IGROV1 was cell-sorted into two subpopulations, P₁–high (66.1%) and P₁-low (33.3%) and we found a significantly higher migration rate in P₁-high expressing cells. Plasma-and ascites-derived natural anti-P₁ IgM bind to the corresponding selfantigen expressed on the ovarian cancer cell surface. These findings are in concordance with the literature on natural IgM as part of the innate immune system recognizing carbo-neo-epitopes. These results deliver the first evidence that P₁ may also be involved in cell migration and therefore may have a role in metastasis.1 page(s

Endogenous galectin-1 enforces class I–restricted TCR functional fate decisions in thymocytes

During thymocyte development, the T-cell receptor (TCR) can discriminate major histocompatibility complex (MHC)/peptide ligands over a narrow range of affinities and translate subtle differences into functional fate decisions. How small differences in TCR input are translated into absolute differences in functional output is unclear. We examined the effects of galectin-1 ablation in the context of class-I–restricted thymocyte development. Galectin-1 expression opposed TCR partial agonist-driven positive selection, but promoted TCR agonist-driven negative selection of conventional CD8+ T cells. Galectin-1 expression also promoted TCR agonist-driven CD8αα intestinal intraepithelial lymphocytes (IEL) development. Recombinant galectin-1 enhanced TCR binding to agonist/MHC complexes and promoted a negative-selection-signaling signature, reflected in intensified rapid and transient extracellular signal-regulated kinase (ERK) activation. In contrast, galectin-1 expression antagonized ERK activity in thymocytes undergoing positive selection. We propose that galectin-1 aids in discriminating TCR-directed fate decisions by promoting TCR binding to agonist/MHC complexes and enforcing agonist-driven signals, while opposing partial-agonist signals. In this way, galectin-1 widens the distinction between TCR-directed functional fate cues