Background: Preparation of highly standardized polyherbal formulation with its chief active chemical constituents supported by therapeutic efficacy in vitro is a valuable approach in the field of pharmaceutical sciences. Objective: The present work aims to develop the high‑performance thin‑layer chromatography (HPTLC) marker‑based standardization of polyherbal formulation using Piperine, Asiaticoside,and Withanolide‑A and in vitro acetylcholinesterase inhibition activity.Materials and Methods: For successful standardization, the HPTLC quantification of Piperine, Asiaticoside, and Withanolide‑A was carried out. Suitable solvent systems were optimized to achieve the better resolution of the marker compounds, extracts, and sample formulation.The reproducibility of the methods was also confirmed by repeating the procedure twice. The identity of the bands in the sample formulation was confirmed by comparing the Rf value with those of their respective reference standards. In vitro ...
Human melanoma cells (M21) actively attach and spread on a fibronectin substrate. Indirect immunofluorescence assays with specific monoclonal antibodies directed to the disialoganglioside GD2, the major ganglioside expressed on M21 melanoma cells, indicate that during the cell attachment process this molecule redistributes into microprocesses that make direct contact with the fibronectin substrate. Scanning and transmission immunoelectron microscopic studies with anti-GD2 monoclonal antibodies and immuno-gold staining demonstrate that GD2 preferentially localizes into substrate-associated microprocesses that emanate from the plasma membrane of the M21 cells. Staining with monoclonal antibodies directed to other melanoma surface antigens fails to demonstrate a similar distribution pattern on these cells. Direct evidence is provided that GD2 is involved in M21 cell attachment to fibronectin, since treatment of these cells with anti-GD2 monoclonal antibodies causes cell rounding and detachment from ...
Background: Anti-ganglioside antibody assays are widely used for diagnosis of autoimmune peripheral neuropathies. Objective: This study aimed to determine serum levels of anti-ganglioside antibodies in children with Guillain-Barre syndrome by immunoblotting technique and compare the results with those obtained by ELISA method. Method: In this investigation, 50 children with Guillain-Barre syndrome (GBS) who were admitted from July 2006 to July 2008, to Tabriz Children's hospital in the northwest of Iran were studied. 30 children admitted for various other reasons than GBS were randomly selected as a control group. The levels of anti-ganglioside antibodies in serum were measured by ELISA and immunoblotting methods using commercial kits. Results: Anti-ganglioside antibodies (IgG) were detected in 16 (32%) GBS patients and in 1 (3.3%) control using ELISA assay. However, by employing immunoblotting technique, antibodies against seven gangliosides were found positive in 28 (56%) GBS patients and none in the
Phospholipid patterns of 15 representative strains of the genus Amycolatopsis were recorded by two-dimensional thin-layer chromatography. The structure analysis of the isolated phospholipids was verified by fast atom bombardment-mass spectroscopy. The positive- and negative-ion spectra of the partially purified phospholipid fractions qualitatively reflect their distinctive composition. All strains contained diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylinositol. Two different types of phosphatidylethanolamine and phosphatidylmethylethanolamine were detected, viz., compounds with or without hydroxy fatty acids. These phospholipid patterns underline the integrity of the genus. Fast atom bombardment-mass spectrometry analysis of phospholipid patterns may serve as an aid for differentiation of bacterial species.
David Frommhold, Andreas Ludwig, M. Gabriele Bixel, Alexander Zarbock, Inna Babushkina, Melitta Weissinger, Sandra Cauwenberghs, Lesley G. Ellies, Jamey D. Marth, Annette G. Beck-Sickinger, Michael Sixt, Bärbel Lange-Sperandio, Alma Zernecke, Ernst Brandt, Christian Weber, Dietmar Vestweber, Klaus Ley, Markus Sperandio ...
Ganglioside GM2 (d18:0/23:0) is a ganglioside. A ganglioside is a compound composed of a glycosphingolipid (ceramide and oligosaccharide) with one or more sialic acids (AKA n-acetylneuraminic acid, NANA) linked on the sugar chain. The 60+ known gangliosides differ mainly in the position and number of NANA residues. It is a component of the cell plasma membrane that modulates cell signal transduction events. It appears that they concentrate in lipid rafts. They have recently been found to be highly important in immunology. Natural and semisynthetic gangliosides are considered possible therapeutics for neurodegenerative disorders. Gangliosides are more complex glycosphingolipids in which oligosaccharide chains containing N-acetylneuraminic acid (NeuNAc) are attached to a ceramide. NeuNAc, an acetylated derivative of the carbohydrate sialic acid, makes the head groups of Gangliosides anionic. NB: the M in GM2 stands for monosialo, i.e., one NeuNAc residue. GM2 is the second monosialo ganglioside ...
A radiometric method for the assay of ganglioside sialidase in cultured human fibroblasts was set up. As substrate, highly radioactive (1.28 Ci/mmol) ganglioside GD1a isotopically tritium-labeled at carbon C-3 of the long chain base was employed; the liberated, and TLC separated [3H]GM1 was determined by computer-assisted radiochromatoscanning. Under experimental conditions that provided a low and quite acceptable (4-5%) coefficient of variation, the detection limit of the method was 0.1 nmol of liberated GM1, using as low as 10 μg of fibroblast homogenate as protein. The detection limit could be lowered to 0.02-0.03 nmol, adopting conditions that, however, carried a higher analytical error (coefficient of variation over 10%). The content of ganglioside sialidase in human fibroblasts cultured in 75-cm2 plastic flasks was 5.8 ∓ 2.5 (SD) nmol liberated GM1 h-1 mg protein-1. Subfractionation studies performed on fibroblast homogenate showed that the ganglioside sialidase was mainly associated ...
Pesticides are poisons in nature and have harmful effects on human health. They are used for suicide, homicide and for other bad intentions. Objective: The aim of the study was to determine the residues of some pesticides in fruits collected from various markets of Lahore. Material & Methods: Eight fruits samples apple, banana, guava, melon, orange, papaya, pomegranate and strawberry from various sale points of the market were collected. Extraction, clean up and analysis were then processed. Concentration of five pesticides including Bifenthrin, Lambda Cyhalothrin, Carbofuran, Chlorpyrifos and Cypermethrin were determined in fruits samples using high performance thin layer chromatography. Spot diameter of pesticide residue of sample was compared with that of standards to determine pesticide concentration. One way ANOVA was applied to determine the significance of difference. Results & Cnoclusion: Significant difference of pesticide presence was identified in some selected fruits ( ...
Fibroblast growth factor receptors (FGFRs) have been implicated in the malignant transformation and chemoresistance of epithelial ovarian cancer; however, the underlying molecular mechanisms are poorly understood. Increased sialyltransferase activity that enhances protein sialylation is an important post‑translational process promoting cancer progression and malignancy. In the present study, α2,6‑sialyltransferase (ST6Gal‑I) overexpression or knockdown cell lines were developed, and FGFR1 was examined to understand the effect of sialylation on migration and drug resistance, and the underlying mechanisms. It was identified that cells with ST6Gal‑I overexpression had increased cell viability and migratory ability upon serum deprivation. Moreover, ST6Gal‑I overexpression cells had strong resistance to paclitaxel, as demonstrated by low growth inhibition rate and cell apoptosis level. A mechanistic study showed that ST6Gal‑I overexpression induced high α2,6‑sialylation of FGFR1 and ...
A murine monoclonal antibody (monoclonal antibody 126) produced against cultured human neuroblastoma cells (LAN-1) was found to be specifically directed to a disialoganglioside (GD2) antigen preferentially expressed on both cell lines and tissues derived from melanoma and neuroblastoma. In enzyme-linked immunosorbent assays, monoclonal antibody 126 failed to react with leukemic and lymphoblastoid cells as well as with a variety of carcinoma and sarcoma cell lines. Immunohistological analysis by the immunoperoxidase technique revealed strong reactivity of monoclonal antibody 126 with frozen and formaldehyde-fixed neuroblastoma and melanoma tissues. Tissues from patients with glioma or with small cell cancer of the lung showed faint staining, whereas those from individuals with sarcoma, lymphoma, and a variety of other neoplasms proved to be negative. Sera of neuroblastoma patients showed significantly elevated GD2 levels compared to normal children (p , 0.001) and children with other tumors (p , ...
A 44-year-old male patient with a diagnosis of Miller Fisher syndrome, Graves disease and central nervous system demyelination is presented. Clinical and laboratory findings supported the diagnosis of Miller Fisher syndrome. On T2-weighted sections of cranial magnetic resonance imaging, many ovoid-shaped, hyperintense lesions in bilateral deep white matter were detected. Magnetic resonance imaging spectroscopy demonstrated low N-acetylaspartate to creatine (NAA/Cr) ratio consistent with demyelination. Combined peripheral nervous system and central nervous system demyelinating processes are rare. In this Miller Fisher syndrome case, an immunological mechanism affecting both the peripheral nervous system and central nervous system with association of Graves disease is discussed in light of the relevant literature. A positive response to intravenous immunoglobulin G treatment was obtained.. Keywords: Miller Fisher syndrome, Graves disease, central nervous system, demyelinating diseases ...
We have generated mouse models of human Tay-Sachs and Sandhoff diseases by targeted disruption of the Hexa (α subunit) or Hexb (β subunit) genes, respectively, encoding lysosomal β-hexosaminidase A (structure, α) and B (structure, ββ). Both mutant mice accumulate GM2 ganglioside in brain, much more so in Hexb −/− mice, and the latter also accumulate glycolipid GA2. Hexa −/− mice suffer no obvious behavioral or neurological deficit, while Hexb −/− mice develop a fatal neurodegenerative disease, with spasticity, muscle weakness, rigidity, tremor and ataxia. The Hexb −/− but not the Hexa −/− mice have massive depletion of spinal cord axons as an apparent consequence of neuronal storage of GM2. We propose that Hexa −/− mice escape disease through partial catabolism of accumulated GM2 via GA2 (asialo-GM2) through the combined action of sialidase and β-hexosaminidase B.. ...
Thin layer chromatography (TLC) with densitometry has been established for the identification and the quantification of inosine pranobex in drug substance and drug products. Inosine pranobex is a combination of inosine, acetamidobenzoic acid, and dimethylaminoisopropanol. UV densitometry was performed in absorbance mode at 260 nm. The separation was carried out on aluminum sheet of silica gel 60 f 254 [chloroform - methanol- - toluene -10% ammonia solution (6:5:1: 0.1% v/v)] as mobile phase. Linearity range was found to be 1-12, 2-12, 2-20 and 2-16 µg/ml for inosine pranobex, inosine, acetamidobenzoic acid, and dimethylaminoisopropanol with the mean percentage recoveries 99.74± 1.73%, 99.88 ± 1.75%, 99.56 ±1.08%, and 99.36 ± 0.71% respectively, (Correlation coefficient r2 = 0.9998 for inosine pranobex, r2 = 0.09999 for inosine, r2 = 0.9998 for acetamidobenzoic acid and r2= 0.9998 for dimethylaminoisopropanol). The detection and quantification limits for inosine pranobex and other components are
TY - JOUR. T1 - A multifunctional Pasteurella multocida sialyltransferase. T2 - A powerful tool for the synthesis of sialoside libraries. AU - Yu, Hai. AU - Chokhawala, Harshal. AU - Karpel, Rebekah. AU - Yu, Hui. AU - Wu, Bingyuan. AU - Zhang, Jianbo. AU - Zhang, Yingxin. AU - Jia, Qiang. AU - Chen, Xi. PY - 2005/12/21. Y1 - 2005/12/21. N2 - A multifunctional sialyltransferase has been cloned from Pasteurella multocida strain P-1059 and expressed in E. coli as a truncated C-terminal His6-tagged recombinant protein (tPm0188Ph). Biochemical studies indicate that the obtained protein is (1) an α2,3-sialyltransferase (main function), (2) an α2,6-sialyltransferase, (3) an α2,3-sialidase, and (4) an α2,3-trans-sialidase. The recombinant tPm0188Ph is a powerful tool in the synthesis of structurally diverse sialoside libraries due to its relaxed substrate specificity, high solubility, high expression level, and multifunctionality.. AB - A multifunctional sialyltransferase has been cloned from ...
Sandhoff disease is a rare, inherited lipid storage disorder that progressively destroys nerve cells in the brain and spinal cord. It is caused by a deficiency of the enzyme beta-hexosaminidase, which results in the harmful accumulation of certain fats (lipids) in the brain and other organs of the body. Sandhoff disease is a severe form of Tay-Sachs disease, the incidence of which had been particularly high in people of Eastern European and Ashkenazi Jewish descent, but Sandhoff disease is not limited to any ethnic group. Each parent must carry the defective gene and pass it on to the child. Individuals who carry only one copy of the mutated gene typically do not show signs and symptoms of the disorder. Onset of the disorder usually occurs at 6 months of age. Symptoms may include:. ...
TY - JOUR. T1 - Gangliosides and neutral glycolipids in ependymal, neuronal and primitive neuroectodermal tumors. AU - Yates, Allan J.. AU - Franklin, Teresa K.. AU - McKinney, Paula. AU - Collins, Raymond. AU - Comas, Theodore. AU - Boesel, Carl P.. AU - Pearl, Dennis K.. N1 - Funding Information: This work was supported by grants from The National Cancer Institute UO1-CA50910, P30 CA 16058, the NCI Cooperative Human Tissue Network, The Department of Pathology and the College of Medicine, and Public Health, OSU.. PY - 1999. Y1 - 1999. N2 - Neutral glycolipid and ganglioside compositions were determined on 11 ependymal tumors, 12 medulloblastomas, 6 other neuronal tumors of the brain, 4 peripheral neuroblastomas, 1 cerebral primitive neuroectodermal tumor (PNET), and 1 PNET of the thoracic wall. Within the group of tumors that can demonstrate neuronal phenotypes, there was an association between the degree of neuronal differentiation usually demonstrated by these tumors and the proportions of ...
TY - JOUR. T1 - Chemical reactions in fast atom bombardment mass spectrometry. AU - Vékey, K.. AU - Zerilli, Luigi F.. PY - 1991. Y1 - 1991. N2 - Examples of various chemical reactions occurring in the matrix or in the selvedge region in fast atom bombardment (FAB) mass spectrometry are discussed. These are categorized as oxidations and reductions; substitutions; clusterings and additions; and sample decomposition or transformation. Some reactions observed showed significant time behaviour and in one case it was possible to determine rate constants. These data suggest that chemical reactions can be accelerated significantly by fast atom bombardment.. AB - Examples of various chemical reactions occurring in the matrix or in the selvedge region in fast atom bombardment (FAB) mass spectrometry are discussed. These are categorized as oxidations and reductions; substitutions; clusterings and additions; and sample decomposition or transformation. Some reactions observed showed significant time ...
TY - JOUR. T1 - Fuc-GM1 ganglioside mimics the receptor function of GM1 for cholera toxin. AU - Masserini, M.. AU - Freire, E.. AU - Palestini, P.. AU - Calappi, E.. AU - Tettamanti, G.. PY - 1992. Y1 - 1992. N2 - The ability of Fuc-GM1 ganglioside to mimic the receptor function of GM1 for cholera toxin (CT) has been investigated. For this purpose, rat glioma C6 cultured cells were enriched with Fuc-GM1 and the responsiveness to CT was compared with that of cells enriched with GM1 ganglioside. Fuc-GM1 was taken up by cells as rapidly and to the same extent as GM1. When comparable amounts of ganglioside were associated, the cells enriched with Fuc-GM1 bound the same amount of 125I-CT as did cells enriched with GM1. Under conditions in which GM1- and Fuc-GM1-enriched cells bound comparable amounts of CT, the Fuc-GMl-treated cells accumulated virtually the same amount of cyclic AMP as did GM1-treated cells, and activation of adenylate cyclase was also similar. The lag time preceding the CT-induced ...
TY - JOUR. T1 - Production of a fusion protein consisting of the enterotoxigenic Escherichia coli heat-labile toxin B subunit and a tuberculosis antigen in Arabidopsis thaliana. AU - Rigano, M. M.. AU - Alvarez, M. L.. AU - Pinkhasov, J.. AU - Jin, Y.. AU - Sala, F.. AU - Arntzen, C. J.. AU - Walmsley, A. M.. PY - 2004/2. Y1 - 2004/2. N2 - Transgenic plants are potentially safe and inexpensive vehicles to produce and mucosally deliver protective antigens. However, the application of this technology is limited by the poor response of the immune system to non-particulate, subunit vaccines. Co-delivery of therapeutic proteins with carrier proteins could increase the effectiveness of the antigen. This paper reports the ability of transgenic Arabidopsis thaliana plants to produce a fusion protein consisting of the B subunit of the Escherichia coli heat-labile enterotoxin and a 6 kDa tuberculosis antigen, the early secretory antigenic target ESAT-6. Both components of the fusion protein were detected ...
Sandhoff disease involves the CNS accumulation of ganglioside GM2 and asialo-GM2 (GA2) due to inherited defects in the beta-subunit gene of beta-hexosaminidase A and B (Hexb gene). Accumulation of these glycosphingolipids (GSLs) produces progressive neurodegeneration, ultimately leading to death. Substrate reduction therapy (SRT) aims to decrease the rate of glycosphingolipid (GSL) biosynthesis to compensate for the impaired rate of catabolism. The imino sugar, N-butyldeoxygalactonojirimycin (NB-DGJ) inhibits the first committed step in GSL biosynthesis. NB-DGJ treatment, administered from postnatal day 2 (p-2) to p-5 (600 mg/kg/day)), significantly reduced total brain ganglioside and GM2 content in the Sandhoff disease (Hexb(-/-)) mice, but did not reduce the content of GA2. We also found that NB-DGJ treatment caused a slight, but significant elevation in brain sialidase activity. The drug had no adverse effects on viability, body weight, brain weight, or brain water content in the mice. No significant
Definition of Miller fisher syndrome with photos and pictures, translations, sample usage, and additional links for more information.
Ganglioside GM1 (18:1/12:0) is a glycosphingolipid (ceramide and oligosaccharide)or oligoglycosylceramide with one or more sialic acids (i.e. n-acetylneuraminic acid) linked on the sugar chain. It is a component the cell plasma membrane which modulates cell signal transduction events. Gangliosides have been found to be highly important in immunology. Ganglioside GM1 carries a net-negative charge at pH 7.0 and is acidic. Gangliosides can amount to 6% of the weight of lipids from brain, but they are found at low levels in all animal tissues. Gangliosides are glycosphingolipids. There are four types of glycosphingolipids, the cerebrosides, sulfatides, globosides and gangliosides. Gangliosides are very similar to globosides except that they also contain N-acetyl neuraminic acid (NANA) in varying amounts. The specific names for the gangliosides provide information about their structure. The letter G refers to ganglioside, and the subscripts M, D, T and Q indicate that the molecule contains mono-, ...
BACKGROUND: GQ1b antibody (GQ1b-Ab) is detected in approximately two-thirds of sera of patients with Bickerstaffs encephalitis (BE). Whilst some of the remaining patients have antibodies to other gangliosides, many patients with BE are reported to be seronegative. METHODS AND RESULTS: Voltage-gated potassium channel antibody (VGKC-Ab) at high titer was detected during the diagnostic work-up of one patient with BE. Sera of an additional patient with BE and nine patients with Miller Fisher syndrome (MF) (all GQ1b-Ab positive) were investigated for VGKC-Ab and other anti-neuronal antibodies by radioimmunoprecipitation using 125I-dendrotoxin-VGKC and immunohistochemistry, respectively. Two patients with MF exhibited moderate titer VGKC-Abs. Regardless of positivity for VGKC or GQ1b antibodies, serum IgG of all patients with BE and MF reacted with the molecular layer and Purkinje cells of the cerebellum in a distinctive pattern. CONCLUSION: Voltage-gated potassium channel antibodies might be involved in some
The structurally similar polysialic acid capsules of group B meningococci and Escherichia coli K1 are poor immunogens, and attempts are currently being made to improve their immunogenicity by chemical modifications. An IgG monoclonal antibody to these polysialic acid capsules was used for the study of the presence of structurally similar components in tissue glycoproteins to investigate the reasons for the poor immunogenicity and to evaluate potential dangers in active or passive immunization. By immunoblotting polysialic acid was detected outside the brain in newborn rat kidney, heart, and muscle. It appeared in immunoblots as one component and with similar mobility to the neural cell adhesion molecule N-CAM. Specificity studies of the antibody and endosialidase treatment showed that the polysialic acid glycans detected were composed of chains as long as eight sialic acid residues or more. The polysialic acid was not detected in the corresponding tissues of the adult animal. These results ...
To determine the hapten number in hapten-carrier protein conjugate matrix-assisted laser desorption/ionization (MALDI) tof mass spectrometry was applied. Highly specific anti-ginsenoside Rb1 and Rg1 monoclonal antibodies (MAbs) were prepared. Ginsenosides were developed on thin layer chromatography (TLC) plates which were covered by a polyvinylidene difluoride (PVDF) membrane resulting in blotting. The membrane was treated with NaIO4 solution to release the aldehyde group on the sugar moiety of the ginsenosides. By treatment of the membrane with a protein solution the ginsenoside-protein conjugation as a Schiff-base occurred, which can function to fix it to the PVDF membrane. A part of the ginsenoside aglycone was reacted with anti-ginsenoside Rb1 MAb, secondary MAb conjugated with enzyme and finally a substrate was added, resulting in a specific and highly sensitive staining that we named Eastern blotting. Furthermore, it makes one-step isolation of ginsenoside Rb1 possible using an immuno-affinity
Autoimmune neuropathies are frequently associated with pathogenic anti-ganglioside antibodies targeting ganglioside-rich neuronal and glial membranes. The extent of injury is determined by the concentration of membrane ganglioside and thus reduction might be expected to attenuate disease. In this study, we suppressed ganglioside biosynthesis in PC12 cells with the glucosylceramide synthase inhibitor, N-butyldeoxynojirimycin and observed reduced plasma membrane antibody binding and a major neuroprotective effect in complement-mediated lysis assays. These data demonstrate that iminosugar inhibitors, currently used to treat type 1 Gaucher disease, are also of potential value for depleting antigen and thereby suppressing tissue injury in anti-ganglioside antibody-associated neuropathy. ...
Tay-Sachs disease is a genetic disorder that results in the destruction of nerve cells in the brain and spinal cord. The most common type, known as infantile Tay-Sachs disease, becomes apparent around three to six months of age with the baby losing the ability to turn over, sit, or crawl. This is then followed by seizures, hearing loss, and inability to move. Death usually occurs in early childhood. Less commonly the disease may occur in later childhood or adulthood. These forms are generally milder in nature. Tay-Sachs disease is caused by a genetic mutation in the HEXA genes on chromosome 15. It is inherited from a person's parents in an autosomal recessive manner. The mutation results in problems with an enzyme called beta-hexosaminidase A which results in the buildup of the molecule GM2 ganglioside within cells, leading to toxicity. Diagnosis is by measuring the blood hexosaminidase A level or genetic testing. It is a type of sphingolipidosis. The treatment of Tay-Sachs disease is supportive ...
Motor neuropathies and multifocal motor neuropathy with conduction block are treatable causes of neuropathy that present with the clinical syndrome of
Beta-hexosaminidase subunit beta is an enzyme that in humans is encoded by the HEXB gene. Hexosaminidase B is the beta subunit of the lysosomal enzyme beta-hexosaminidase that, together with the cofactor GM2 activator protein, catalyzes the degradation of the ganglioside GM2, and other molecules containing terminal N-acetyl hexosamines. Beta-hexosaminidase is composed of two subunits, alpha and beta, which are encoded by separate genes. Both beta-hexosaminidase alpha and beta subunits are members of family 20 of glycosyl hydrolases. Mutations in the alpha or beta subunit genes lead to an accumulation of GM2 ganglioside in neurons and neurodegenerative disorders termed the GM2 gangliosidoses. Beta subunit gene mutations lead to Sandhoff disease (GM2-gangliosidosis type II). The HEXB gene lies on the chromosome location of 5q13.3 and consists of 15 exons, spanning 35-40Kb. HEXB consists of 556 amino acid residues and weighs 63111Da. HEXB is one of the two subunits forming β-hexosaminidase which ...
Anti-idiotype vaccination represents an innovative approach to target tumor-associated antigen-expressing cells. This approach comes directly from Jerne's idiotypic network theory, which postulates that due to the huge potentiality for diversity of the immunoglobulin variable regions, the idiotype repertoire can mimic the universe of self and foreign epitopes [1].. NeuGc-containing gangliosides are attractive targets for cancer immunotherapy because these glycolipids are non-self antigens in humans [2, 3]. In contrast, they have been detected in different human tumors by antibodies and chemical analysis [4-6]. Recent experimental data suggest that N-glycolyl-GM3 ganglioside (NeuGcGM3) is relevant for tumor biology [7].. mAb-1E10 [8] is an IgG1 anti-idiotype (Ab2) mAb obtained by immunizing Balb/c mice with mAb-P3 (Ab1) [9] coupled to keyhole limpet hemocyanin (KLH) in the presence of Freund's adjuvant. This Ab2 inhibited the binding of mAb-P3 to NeuGcGM3 ganglioside. mAb-1E10 induced an ...
Franklin for consultation services on the neuropathology and for making plates from his forthcoming A Stereotaxic Atlas of the Mouse Brain (K. Franklin and G. Paxinos, eds, Academic Press) accessible prior to publication, Mirielle. Shevell for help with the particular phenotype, L. Old for helping make the ganglioside analysis possible, B.. T, testis; E, epididymis; Lu, chest; Li, liver; K, renal; H, heart; B, mind; S, spleen. ES tissues in 800 µl associated with cold PBS using a Bio-Rad GenePulser (240V, five-hundred µF).. (C) plus (F) are from outdoors type, (A), (D) plus (G) from Hexa −/− and (B), (E) and (H) from Hexb −/− mice. (A) and (B) Electron micrographs of cortical neurons showing large clusters of MCB. Some lysosomes have membrane whorls.. In the particular kidney, the cytoplasm associated with epithelial cells lining the particular proximal tubules showed considerable vacuolation (Fig. 7H). Renal corpuscles, distal convoluted tubules and collecting ducts came out normal. ...
Fuc-GM1 is extensively expressed on most SCLCs and minimally expressed on normal tissues, suggesting that this ganglioside antigen may serve as an excellent target for active immunization. However, the immunogenicity of autoantigens such as Fuc-GM1 cannot be consistently predicted based on expression in normal cells. The GD3 ganglioside, which has a restricted distribution on normal cells that is limited to the brain, connective tissue, and a small population of T cells, has proven to be poorly immunogenic in humans (30 , 31) , although occasional antibody responses against GD3 have been induced (32) . In contrast, GM2, which is expressed in the brain as well as the secretory borders of all epithelial tissues, has proven to be highly immunogenic. Fuc-GM1 has a more restricted distribution on normal tissues than either GM2 or GD3 and, therefore, would be expected to be more immunogenic. This study demonstrates that indeed this is the case.. Median peak ELISA antibody titers against Fuc-GM1 after ...
Diarrhoea caused by enterotoxigenic Escherichia coli is one of the leading causes of mortality in children under five years of age and is a great burden on developing countries. The major virulence factor of the bacterium is the heat-labile enterotoxin (LT), a close homologue of the cholera toxin. The toxins bind to carbohydrate receptors in the gastrointestinal tract, leading to toxin uptake and, ultimately, to severe diarrhoea. Previously, LT from human- and porcine-infecting ETEC (hLT and pLT, respectively) were shown to have different carbohydrate-binding specificities, in particular with respect to N-acetyllactosamine-terminating glycosphingolipids. Here, we probed eleven single-residue variants of the heat-labile enterotoxin with surface plasmon resonance spectroscopy and compared the data to the parent toxins. In addition we present a 1.45 Å crystal structure of pLTB in complex with branched Lacto-N-neohexaose (Galbeta4GlcNAcbeta6[Galbeta4GlcNAcbeta3]Galbeta4Glc). The largest difference in
The post infectious paralytic autoimmune disease, Guillain-Barré syndrome (GBS), has been associated with the generation of cross-reactive auto-antibodies after Campylobacter jejuni infection. These auto-antibodies interact with both the ganglioside mimicking C. jejuni lipo-oligosaccharides (LOS) and endogenous gangliosides. This study sought to investigate novel interactions of the ganglioside mimicking LOS with immune system ganglioside specific receptors. In addition, studies investigated if such receptor recognition affects antigen trafficking or the immunostimulatory potency of the LOS, which could participate in auto-antibody generation. Results presented in this thesis demonstrate for the first time that certain members of the siglec receptor family are capable of recognising LOS from a GBS associated strain of C. jejuni. This interaction did not definitively result in enhanced, or altered, potency of ganglioside mimicking LOS in stimulating immune cells. Interestingly, ganglioside ...
This invention provides a novel ganglioside ceramidase which has a substrate specificity at least for GD1a, GM1, GM2 and GM3 and acts at least on GD1a, GM1, GM2 and GM3 and catalyzes the reaction of hydrolysis of ganglioside to lysoganglioside and fatty acid.This invention further provides a process for producing the novel ganglioside ceramidase which comprises cultivating ganglioside ceramidase producing strain belonging to the genus Nocardia in a culture medium and collecting ganglioside ceramidase from the culture.
How is lecithin/sphingomyelin ratios abbreviated? L/S stands for lecithin/sphingomyelin ratios. L/S is defined as lecithin/sphingomyelin ratios rarely.
Fingerprint Dive into the research topics of 'Isolation, purification and characterization of a trypsin from the pyloric ceca of mullet (Mugil cephalus)'. Together they form a unique fingerprint. ...
Title:L-Pyrrolidine-2-Carboxylic Acid-4-Hydrogen Sulfate (Supported on Silica Gel) as a New and Efficient Catalyst for Acylation of Alcohols, Phenols and Amines Under Solvent-Free Conditions. VOLUME: 10 ISSUE: 5. Author(s):Maryam Hajjami, Arash Ghorbani-Choghamarani and Zahra Khani. Affiliation:Department of Chemistry, Faculty of Science, Ilam University, P.O. Box 69315516, Ilam, Iran.. Keywords:Acylation, L-pyrrolidine-2-carboxylic acid-4-hydrogen sulfate, Acetic anhydride, Alcohol, Phenol, Amine.. Abstract:In the present work, L-pyrrolidine-2-carboxylic acid-4-hydrogen sulfate, supported on silica gel was prepared and characterized by Mass spectroscopy, 1H NMR, 13CNMR, FT IR and elemental analysis (CHN) methods. This heterogenized catalyst can be used as an efficient catalyst for the acylation of alcohols, phenols, and amines with acetic anhydride under mild and solvent-free conditions. Simple work-up, stability of the catalyst, nontoxicity and good to high yields are the advantages of this ...
article{9a3fcc76-7ff7-4e06-9659-b11bead8e138, abstract = {A 52-year-old man developed diplopia, a nasal voice, dysphagia, hoarseness and slight bilateral facial palsies. There was no ataxia, areflexia, limb weakness or sensory involvement. Serum anti-GQ1b IgG antibody was present. Treatment with intravenous immunoglobulin started, and the patient responded with a rapid resolution of symptoms. The diagnosis is consistent with polyneuritis cranialis which is considered to be a Guillain-Barre syndrome variant, a forme fruste, but very rare. The diagnosis can be difficult and a thorough investigation is required. Electrophysiological examination, laboratory evaluations, imaging and cerebrospinal fluid examination are often required in the investigations. Cranial neuropathy can be the presentation of many disorders. Determination of anti-ganglioside antibodies as anti-GQ1b is valuable to the diagnosis, and shows the association with the Guillain-Barre syndrome.}, author = {Edvardsson, Bengt and ...
Molecular mimicry of Campylobacter jejuni lipo-oligosaccharides (LOS) with gangliosides in nervous tissue is considered to induce cross-reactive antibodies that lead to Guillain-Barre syndrome (GBS), an acute polyneuropathy. To determine whether specific bacterial genes are crucial for the biosynthe …
Radix Rehmanniae is one of beneficial traditional Chinese medicine.Study on the literatures of iridoid glucosides from Radix Rehmanniae shows that Catalpol has hypoglycemic effect on senile dementia and Parkinson disease.Iridoid glucosides from Radix Rehmanniae have large polarity,and it is difficult to be isolated from their similar structures.Combined the nature of catalpol with the study of literature,the following extraction process was taken: After the recirculation of Rehmanniae glutinosa by acetone,most of saccharide in acetone extracts is gotten rid of by activated carbon column chromatography,and then by silica gel column chromatography,to be crude,finally recrystallized to be catalpol.Then the thin layer chromatography(TLC) and high performance liquid chromatography(HPLC) method were used to determine the catalpol.The purity is 98% while 0.25 μg Catalpol was tested in HPLC.Rf figures and colors of Catalpol are the same with those of standard samples in TLC.Finally,the acute toxicity test of
TY - JOUR. T1 - The role of ganglioside GM3 in the modulation of conformation and activity of sarcoplasmic reticulum CA2+-ATPase. AU - Yang, F. Y.. AU - Wang, L. H.. AU - Yang, X. Y.. AU - Tsui, Z. C.. AU - Tu, Yaping. PY - 1997/10. Y1 - 1997/10. N2 - Rabbit sarcoplasmic reticulum does contain trace amounts of gangliosides, and the main species is GM3. Incorporation of GM3 into the SR vesicles or addition of it to the soybean phospholipid used for reconstitution of proteoliposomes obviously increased ATP hydrolysis, as well as, Ca2+ uptake activity of sarcoplasmic reticulum Ca2+-ATPase. Conformation changes of Ca2+-ATPase induced by GM3 were also observed by circular dichroism, intrinsic fluorescence and fluorescence quenching measurements.. AB - Rabbit sarcoplasmic reticulum does contain trace amounts of gangliosides, and the main species is GM3. Incorporation of GM3 into the SR vesicles or addition of it to the soybean phospholipid used for reconstitution of proteoliposomes obviously increased ...
Other Development of an effective low-cost anti-acquired immunodeficiency syndrome (AIDS) drugs is needed for treatment of AIDS patients in developing countries. Host cell lipid raft microdomains, which are enriched with cholesterol, glycolipids, ceramide, and gangliosides, are important for human immunodeficiency virus type 1 (HIV-1) entry. Retinoid analogs have been shown to modulate ceramide levels in the cell membrane, while cholera toxin B subunit (CT-B) specifically binds to the ganglioside GM1. In this study, we found that the acyclic retinoid analogs geranylgeranoic acid (GGA) and NIK-333 as well as CT-B efficiently attenuate CXCR4-tropic, but not CCR5-tropic, HIV-1 vector infection. We also found that GGA and NIK-333 suppress CXCR4-tropic HIV-1 infection by attenuating CXCR4 expression. CT-B also attenuated CXCR4-tropic HIV-1 infection, but did not suppress CXCR4 expression. These results suggest a distinct role for lipid raft microdomains in CXCR4- and CCR5-tropic HIV-1 infections and ...
Sialic acids are prominent termini of mammalian glycoconjugates and are key binding determinants for cell-cell recog-nition lectins. Binding of the sialic acid-dependent lectin, myelin-associated glycoprotein (MAG), to nerve cells is implicated in the inhibition of nerve regeneration after injury. Therefore, blocking MAG binding to nerve cell sialoglycoconjugates might enhance nerve regeneration. Previously, we reported that certain sialoglycoconjugates bearing N-acetylneuraminic acid (NeuAc) but not N-glycolylneuraminic acid (NeuGc) support MAG binding (Collins et al., 1997a). We now report highly efficient conversion of sialic acids on living neural cells from exclusively NeuAc to predominantly NeuGc using a novel synthetic metabolic precursor, N-glycolylmannosamine pentaacetate (Man-NGc-PA). When NG108-15 neuroblastoma-glioma hybrid cells, which normally express only NeuAc (and bind to MAG), were cultured in the presence of 1 mM ManNGcPA, they expressed 80-90% of their sialic acid precursor pool as
The mechanism of transsulfuration and the role of cystathionine in the biosynthesis of methionine in yeast were investigated. Saccharomyces were shown to accumulate cystathionine by use of the ³⁵S labeled compound. Cell-free extracts of the wildtype clone as well as methionine auxotrophs were shown to cleave cystathionine. The enzymatically produced compounds were shown to be pyruvic acid and homocysteine. Pyruvic acid was determined by the Rf value of the 2, 4-dinitrophenylhydrazone as compared to authentic. The absorption spectrum of the phenylhydrazone of authentic pyruvate was identical to that of the phenylhydrazone of the enzymatic reaction mixture. The sulfhydryl was shown to be homocysteine by comparing the Rf values of the radioactive cleavage product to a ninhydrin and sulfur positive reference spot of homocysteine, and by bioautographic procedures. Transsulfuration and the role of cystathionine in methionine biosynthesis in yeast were discussed in view of these results which varied ...
The study is designed to further examine the extent to which GM1 ganglioside can improve symptoms, delay disease progression, and, perhaps, partially restore damaged brain cells in PD patients. GM1 ganglioside is a chemical that is normally found in the brain and is a normal part of the outer covering or membrane of nerve cells. This study will compare the effectiveness of GM1 to standard PD treatment. In addition to studying clinical measures of motor and cognitive functioning, the investigators will use PET (positron emission tomography) scanning to image the brain and the dopamine nerve endings in a subgroup of patients. Patients with mild to moderate idiopathic PD will be divided into 2 groups. One group will receive GM1 for 24 weeks and the other will receive placebo. At the end of this 24 week period, all patients will enter into a 96 week treatment period in which all patients will receive GM1.. In parallel, a group of standard-of-care patients with mild to moderate PD will be monitored ...
A deficiency of lysosomal β-d-galactosidase (βG; EC 3.2.1.23) is the primary defect in the three clinical forms (infantile, juvenile, and adult) of GM1 gangliosidosis and in Morquio B syndrome. Patients with the infantile form of GM1 gangliosidosis (type 1), who usually die before the age of 3 years, display the coarse face, hepatosplenomegaly, and skeletal dysplasia reminiscent of Hurler disease. Cases with later onset, described as the late infantile/juvenile form (type 2), display progressive psychomotor loss but less prominent dysmorphic changes. Extrapyramidal signs of protracted course are the major neurologic manifestations in the adult/chronic form (type 3) of GM1 gangliosidosis. Morquio B syndrome is expressed as generalized skeletal dysplasia with corneal clouding and normal intelligence (1).. βG activity is also severely decreased in galactosialidosis, a disorder resulting from deficiency in cathepsin A/protective protein. This lysosomal protein forms a complex with βG and a ...
We determined the ability of mixed gangliosides (16% GD1b, 19% GT1b, 21% GM1, and 40% GD1a) and individual gangliosides GM1 and GD1b to modulate the NV-PLA2 induced human erythrocyte ghost membrane damage. CM-Sephadex purification of crude Naja naja venom yielded eight peaks of which peak VII, a major phospholipase A2 (NV-PLA2) accounted for 22% of the total protein recovered and 8% of the total PLA2 activity recovered. The membrane damage induced by NV-PLA2 was assessed by measuring the decrease in the relative intensity of fluorescence using cis-parinaric acid (PnA) as a monitor molecule. The RBC membranes isolated from healthy human blood showed 72% damage on treatment with NV-PLA2 (2 mg) when compared to untreated membranes. Mixed gangliosides (18 nM) and GM1 (15 nM) offered 81 and 86% protection respectively, whereas GD1b (20 nM) did not show significant protection. Analysis of membrane bound Na+K+ and Ca2+Mg2+ ATPase indicated a 3 fold and 2 folds decrease in their activities on NV-PLA2
A novel agglutinin with specificity for sialic acid sequence of sugars in thyroglobulin is identified in the hemolymph of Scylla serrata. The physico-chemical characteristics of its binding affinity,...
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