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Gal
Terms modified by Gal Selected AbstractsA hVIPR transgene as a novel tool for the analysis of circadian function in the mouse suprachiasmatic nucleusEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 11 2003V. M. King Abstract A mouse bearing a novel transgene encoding the human VPAC2 receptor (hVIPR; Shen et al. (2000) PNAS, 97, 11575,11580) was used to investigate circadian function in the hypothalamic suprachiasmatic nuclei (SCN). Neurons expressing hVPAC2R, detected by a beta-galactosidase (,-GAL) tag, have a distinct distribution within the SCN, closely matching that of neurophysin (NP) neurons and extending into the region of peptide histidine isoleucine (PHI) cells. In common with NP and PHI cells, neurons expressing hVPAC2R are circadian in nature, as revealed by synchronous rhythmic expression of mPERIOD (mPER) proteins. A population of SCN cells not expressing PHI, NP or hVPAC2R exhibited circadian PER expression antiphasic with the rest of the SCN. Nocturnal light exposure induced mPER1 in the ventral SCN and mPER2 widely across the nucleus. Induction of nuclear mPER2 in hVPAC2R cells confirmed their photic responsiveness. Having established their circadian properties, we tested the utility of SCN neurons expressing the hVIPR transgene as functionally and anatomically explicit markers for SCN tissue grafts. Prenatal SCN tissue from hVIPR transgenic pups survived transplantation into adult CD1 mice, and expressed ,-GAL, PER and PHI. Over a series of studies, hVIPR transgenic SCN grafts restored circadian activity rhythms to 17 of 72 arrhythmic SCN lesioned recipients (23.6%). By using heterozygous hVIPR transgenic grafts on a heterozygous Clock mutant background we confirmed that restored activity rhythms were conferred by the donor tissue. We conclude that the hVIPR transgene is a powerful and flexible tool for examination of circadian function in the mouse SCN. [source] Pathophysiological significance of senescence marker protein-30GERIATRICS & GERONTOLOGY INTERNATIONAL, Issue 2010Naoki Maruyama A novel rat liver protein of 30 kDa, SMP30 decreases with aging. This protein is expressed most prominently in the liver and kidneys among the various organs. Its gene is located on the X chromosome. No functional domain was recognized in the entire amino acid sequence. Recently, we found a homology between rat SMP30 and two species of bacterial gluconolactonase (EC 3.1.1.17). The lactonase reaction with l -gulono-,-lactone is the penultimate step in vitamin C (l -ascorbic acid) biosynthesis. SMP30-knockout (KO) mice fed a vitamin C-deficient diet displayed symptoms of scurvy. In SMP30-KO mice, hepatocytes were more susceptible to apoptosis induced by TNF-, plus actinomycin D than hepatocytes from wild-type mice. Two morphological features considered to be a hallmark of senescence are apparent in SMP30-KO mice. At 12 months of age, SMP30-knockout mice had clearly visible deposits of lipofuscin and senescence-associated ,-galactosidase (SA-,-GAL) in their renal tubular epithelia. These features are compatible with high electron dense deposits in lysosomes. This observation suggests that the SMP30-knockout mouse is a useful model of ordinal senescence. Geriatr Gerontol Int 2010; 10 (Suppl. 1): S88,S98. [source] ACTIVITIES OF ,-GALACTOSIDASE AND ,-L-ARABINOFURANOSIDASE, ETHYLENE BIOSYNTHETIC ENZYMES DURING PEACH RIPENING AND SOFTENINGJOURNAL OF FOOD PROCESSING AND PRESERVATION, Issue 5 2006CHANG-HAI JIN ABSTRACT A study was conducted to determine changes in firmness, ethylene and ethylene biosynthetic enzymes, and the activities of ,-galactosidase (,-GAL) and ,-L-arabinofuranosidase (,-AF) during peach ripening and softening. The activities of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase, ACC oxidase and polygalacturonase increased in parallel with ethylene production and declined in firmness during peach ripening, and they appeared at maximum simultaneously at maturity IV. ,-GAL activity was high in unripe peach fruit and it experienced an overall decline during peach ripening. While ,-AF activity changed placidly at the initial stage (maturity I,III), after that it experienced a rapid increasing stage. The preliminary result indicated that ,-GAL and ,-AF, as well as ethylene biosynthetic enzymes, may be involved in the ripening and softening of peach fruit. [source] Increased Caloric Intake on a Fat-Rich Diet: Role of Ovarian Steroids and Galanin in the Medial Preoptic and Paraventricular Nuclei and Anterior Pituitary of Female RatsJOURNAL OF NEUROENDOCRINOLOGY, Issue 10 2007S. F. Leibowitz Previous studies in male rats have demonstrated that the orexigenic peptide galanin (GAL), in neurones of the anterior parvocellular region of the paraventricular nucleus (aPVN) projecting to the median eminence (ME), is stimulated by consumption of a high-fat diet and may have a role in the hyperphagia induced by fat. In addition to confirming this relationship in female rats and distinguishing the aPVN-ME from other hypothalamic areas, the present study identified two additional extra-hypothalamic sites where GAL is stimulated by dietary fat in females but not males. These sites were the medial preoptic nucleus (MPN), located immediately rostral to the aPVN, and the anterior pituitary (AP). The involvement of ovarian steroids, oestradiol (E2) and progesterone (PROG), in this phenomenon was suggested by an observed increase in circulating levels of these hormones and GAL in MPN and AP with fat consumption and an attenuation of this effect on GAL in ovariectomised (OVX) rats. Furthermore, in the same four areas affected by dietary fat, levels of GAL mRNA and peptide immunoreactivity were stimulated by E2 and further by PROG replacement in E2 -primed OVX rats and were higher in females compared to males. Because both GAL and PROG stimulate feeding, their increase on a fat-rich diet may have functional consequences in females, possibly contributing to the increased caloric intake induced by dietary fat. This is supported by the findings that PROG administration in E2 -primed OVX rats reverses the inhibitory effect of E2 on total caloric intake while increasing voluntary fat ingestion, and that female rats with higher GAL exhibit increased preference for fat compared to males. Thus, ovarian steroids may function together with GAL in a neurocircuit, involving the MPN, aPVN, ME and AP, which coordinate feeding behaviour with reproductive function to promote consumption of a fat-rich diet at times of increased energy demand. [source] Introduction of lipidization,cationization motifs affords systemically bioavailable neuropeptide Y and neurotensin analogs with anticonvulsant activitiesJOURNAL OF PEPTIDE SCIENCE, Issue 9 2010Brad R. Green Abstract The neuropeptides galanin (GAL), neuropeptide Y (NPY) or neurotensin (NT) exhibit anticonvulsant activities mediated by their respective receptors in the brain. To transform these peptides into potential neurotherapeutics, their systemic bioavailability and metabolic stability must be improved. Our recent studies with GAL analogs suggested that an introduction of lipoamino acids in the context of oligo-Lys residues (lipidization,cationization motif) significantly increases their penetration into the brain, yielding potent antiepileptic compounds. Here, we describe an extension of this strategy to NPY and NT. Rationally designed analogs of NPY and NT containing the lipidization,cationization motif were chemically synthesized and their physicochemical and pharmacological properties were characterized. The analogs NPY-BBB2 and NT-BBB1 exhibited increased serum stability, possessed log D > 1.1, retained high affinities toward their native receptors and produced potent antiseizure activities in animal models of epilepsy following intraperitoneal administration. Our results suggest that the combination of lipidization and cationization may be an effective strategy for improving systemic bioavailability and metabolic stability of various neuroactive peptides. Copyright © 2010 European Peptide Society and John Wiley & Sons, Ltd. [source] Galanin Knockout Mice Show Disturbances in Ethanol Consumption and Expression of Hypothalamic Peptides That Stimulate Ethanol IntakeALCOHOLISM, Issue 1 2010Olga Karatayev Background:, There is growing evidence suggesting that hypothalamic galanin (GAL), which is known to stimulate intake of a fat-rich diet, has a role in promoting the consumption of ethanol. The present study further examined this possibility in GAL knockout (GALKO) mice. Methods:, Two groups of female and male GALKO mice, compared to wild-type (WT) controls, were trained to voluntarily drink increasing concentrations of ethanol, while maintained on lab chow and water. They were examined in terms of their daily ethanol intake and preference, acute consumption of a high-fat diet, preference for flavored solutions, and expression of different peptides shown to stimulate ethanol intake. Results:, In the GALKO mice compared to WT, the results revealed: (i) a 35 to 45% decrease in ethanol intake and preference, which was evident only at the highest (15%) ethanol concentration, was stronger in female than in male mice, and was seen with comparisons to littermate as well as nonlittermate WT mice; (ii) a 48% decrease in acute intake of a fat-rich diet, again stronger in female than male mice; (iii) no difference in consumption of sucrose or quinine solutions in preference tests; (iv) a total loss of GAL mRNA in the hypothalamic paraventricular nucleus (PVN) of female and male mice; and (v) a gender-specific change in mRNA levels of peptides in the perifornical lateral hypothalamus (PFLH), orexin and melanin-concentrating hormone, which are known to stimulate ethanol and food intake and were markedly decreased in females while increased in males. Conclusions:, These results provide strong support for a physiological role of PVN GAL in stimulating the consumption of ethanol, as well as a fat-rich diet. Ablation of the GAL gene produced a behavioral phenotype, particularly in females, which may reflect the functional relationship of galanin to ovarian steroids. It also altered the peptides in the PFLH, with their reduced expression contributing to the larger behavioral effects observed in females and their increased expression attenuating these effects in males. [source] Effect of Ethanol on Hypothalamic Opioid Peptides, Enkephalin, and Dynorphin: Relationship With Circulating TriglyceridesALCOHOLISM, Issue 2 2007Guo-Qing Chang Background: Recent evidence has demonstrated that ethanol intake can stimulate the expression and production of the feeding-stimulatory peptide, galanin (GAL), in the hypothalamic paraventricular nucleus (PVN), and that PVN injection of this peptide, in turn, can increase the consumption of ethanol. To test the hypothesis that other feeding-related systems are involved in ethanol intake, this study examined the effect of ethanol on the hypothalamic opioid peptides, enkephalin (ENK), and dynorphin (DYN). Method: Adult, male Sprague,Dawley rats were trained to voluntarily drink increasing concentrations of ethanol, up to 9% v/v, on a 12-hour access schedule or were given a single injection of ethanol (10% v/v) versus saline vehicle. The effect of ethanol on GAL, ENK, and DYN mRNA was measured using real-time quantitative polymerase chain reaction and radiolabeled in situ hybridization, while radioimmunoassay was used to measure peptide levels. In addition to blood alcohol, circulating levels of triglycerides (TG), leptin, and insulin were also measured. Results: The data demonstrated that: (1) rats voluntarily drinking 9% v/v ethanol (approximately 2.0 g/kg/d) show a significant increase in GAL, ENK, and DYN mRNA in the PVN compared with water-drinking rats; (2) voluntary consumption of ethanol also increases peptide levels of ENK and DYN in the PVN; (3) acute injection of 10% ethanol (1.0 g/kg of 10% v/v) similarly increases the expression of GAL, ENK, and DYN in the PVN; and (4) ethanol consumption and injection, while having little effect on leptin and insulin, consistently increase circulating levels of TG as well as alcohol, both of which are strongly, positively correlated with peptide expression in the PVN. Conclusions: These findings, together with published studies, suggest a possible role for hypothalamic opioid peptides in the drinking of ethanol. Based on evidence that dietary fat and lipid injections stimulate the PVN peptides and injection of the opiates and GAL increase ethanol intake, it is proposed that both TG and alcohol in the circulation, which are elevated by the ingestion or injection of ethanol, are involved in stimulating these peptides in the PVN, which in turn promote further consumption of ethanol. [source] Determination of galanthamine in Bulbus Lycoridis Radiatae by coupling capillary electrophoresis with end-column electrochemiluminescence detectionJOURNAL OF SEPARATION SCIENCE, JSS, Issue 15 2010Biyang Deng Abstract A novel method for the determination of galanthamine (GAL) in Bulbus Lycoridis Radiatae has been developed based on coupling CE with an end-column tris(2,2,-bipyridyl)ruthenium(II) electrochemiluminescence (ECL). Parameters affecting CE separation and ECL detection were investigated and optimized. Baseline separation of GAL from other components in the Bulbus Lycoridis Radiatae sample was achieved with an 18,mmol/L phosphate running buffer at pH 9.0. Under the optimized conditions: 12,kV CE-separation voltage, ECL detection potential at 1.25,V with 5,mmol/L and 50,mmol/L phosphate buffer at pH 7.5 in the detection reservoir, the linear range of GAL concentration was from 0.8,ng/mL to 2,,g/mL, whereas the detection limit was 0.25,ng/mL (S/N=3). The proposed method was successfully demonstrated for the determination of GAL in Bulbus Lycoridis Radiatae. [source] Origin and Chemical Coding of Primary Afferent Neurones Supplying the Prostate of the DogANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 6 2004M. B. Arciszewski Summary Retrograde tracing technique combined with the double-fluorescent immunohistochemistry were used to investigate the distribution and chemical coding of primary afferent neurones supplying the canine prostate. After the injection of Fast Blue (FB) into the prostatic tissue retrogradely-labelled (FB+) primary afferent neurones were localized in bilateral L1,Ca1 dorsal root ganglia (DRG). Statistical analysis using anova test showed that there are two major sources of afferent prostate innervation. The vast majority of prostate-supplying primary afferent neurones were located in bilateral L2,L4 DRG (56.9 ± 0.6%). The second source of the afferent innervation of canine prostate were bilateral S1,Ca1 DRG (40.6 ± 1.0%). No statistically significant differences were found between average number of FB+ neurones localized in the left and right DRG (49.5 ± 1.7 and 50.5 ± 1.7%, respectively). Immunohistochemistry revealed that FB+ primary afferent neurones contain several neuropeptides in various combinations. In the prostate-supplying neurones of lumbar and sacro-caudal DRG the immunoreactivity to substance P (SP) and calcitonin gene-related peptide (CGRP) was found most frequently (50 ± 3.7 and 37.3 ± 1.9%, respectively). Both in the lumbar and sacro-caudal DRG, considerable population of FB+ neurones immunoreactive neither to SP nor CGRP were also found (23 ± 2.6 and 32.8 ± 2.3%, respectively). In the lumbar DRG 10.7 ± 1.1% of SP-immunoreactive FB+ neurones also contained galanin (GAL). In 9.2 ± 2.2% of the prostate-supplying primary afferent neurones located in the sacro-caudal DRG the co-localization of SP and GAL was also reported. Results of the retrograde tracing experiment demonstrated for the first time sources of afferent innervation of the canine prostate. Double immunohistochemistry revealed that many of the prostate-supplying primary afferent neurones express some of sensory neuropeptides which presumably may be involved in nociception and some pathological processes like inflammation or nerve injury. [source] Porcine BAC derived microsatellites linked to ADRBK1, CNTF and GAL on SSC2ANIMAL GENETICS, Issue 1 2002M. Faivre No abstract is available for this article. [source] Possible biphasic changes of free radicals in ethylene glycol-induced nephrolithiasis in ratsBJU INTERNATIONAL, Issue 9 2000H.S. Huang Objective To evaluate the possible role of free radicals in nephrolithiasis in rats induced by ethylene glycol, and to examine the correlation between the urinary enzymes N-acetyl-,-glucosaminidase (NAG), ,-galactosidase (GAL) and neutral endopeptidase (NEP), and free radical production. Materials and methods Hyperoxaluria was produced in male Wistar rats by adding ethylene glycol to their drinking water. After 7, 21 and 42 days of treatment, urinary oxalate, creatinine clearance and urinary enzymes (NAG, GAL and NEP) were measured. The nitroblue tetrazolium perfusion method was used to locate the sites of free-radical production. Ultrasensitive chemiluminescence was used to directly measure the production of reactive oxygen species (ROS) in vivo. Vitamin E and potassium citrate were fed to rats, in addition to ethylene glycol, to assess their effects on free radical production. Results Urinary oxalate increased significantly and was associated with an increase in NAG and GAL at all sample times. However, urinary NEP activity was unchanged on day 7, although there was four times as much NEP on days 21 and 42 than in the control groups. Formazan particles in the renal cortex were scored as 3+ to 4+ in rats treated for 7 days with ethylene glycol. Blood ROS levels were also higher in this group than in the controls (P < 0.01). After vitamin E and potassium citrate treatment, blood ROS levels were lower than in rats treated with ethylene glycol alone. Conclusion Free radicals may be produced in the early stages of nephrolithiasis in rats fed with ethylene glycol. Free radicals occurred mainly in blood and might be associated with NEP inactivation. [source] Left-asymmetric expression of Galanin in the linear heart tube of the mouse embryo is independent of the nodal co-receptor gene crypticDEVELOPMENTAL DYNAMICS, Issue 12 2008Axel Schweickert Abstract Only very few left/right asymmetrically expressed genes are known in the mammalian embryo. In a screen for novel factors we identified the gene encoding the neuropeptide Galanin in mouse. At embryonic day (E) 8.5 asymmetric mRNA transcription was found in the left half of the linear heart tube. During heart looping and morphogenesis expression became restricted to the atrio-ventricular (AV) canal, followed by specific staining of the AV-node and AV-rings in the four-chambered heart. Expression was inverted in inv/inv and randomized in homozygous iv mutant embryos. Left-sided heart-specific transcription of mouse Gal thus should be controlled by the left-right pathway. The asymmetric pattern was retained in cryptic mutant embryos, in which the Nodal signaling cascade is disrupted. Surprisingly, Pitx2c was found to be expressed in 50% of cryptic mutant hearts as well, suggesting that some aspects of asymmetric gene expression in the heart are independent of cryptic. Developmental Dynamics 237:3557,3564, 2008. © 2008 Wiley-Liss, Inc. [source] A Lactulose Bienzyme Biosensor Based on Self-Assembled Monolayer Modified ElectrodesELECTROANALYSIS, Issue 17 2004Susana Campuzano Abstract A bienzyme biosensor in which the enzymes ,-galactosidase (,-Gal), fructose dehydrogenase (FDH), and the mediator tetrathiafulvalene (TTF) were coimmobilized by cross-linking with glutaraldehyde atop a 3-mercaptopropionic acid (MPA) self-assembled monolayer on a gold disk electrode, is reported. The working conditions selected were Eapp=+0.10,V and (25±1),°C. The useful lifetime of one single TTF-,-Gal-FDH-MPA-AuE was surprisingly long, 81,days. A linear calibration plot was obtained for lactulose over the 3.0×10,5,1.0×10,3,mol L,1 concentration range, with a limit of detection of 9.6×10,6,mol L,1. The effect of potential interferents (lactose, glucose, galactose, sucrose, and ascorbic acid) on the biosensor response was evaluated. The behavior of the SAM-based biosensor in flow-injection systems in connection with amperometric detection was tested. The analytical usefulness of the biosensor was evaluated by determining lactulose in a pharmaceutical preparation containing a high lactulose concentration, and in different types of milk. Finally, the analytical characteristics of the TTF-,-Gal-FDH-MPA-AuE are critically compared with those reported for other recent enzymatic determinations of lactulose. [source] Casein phosphoproteome: Identification of phosphoproteins by combined mass spectrometry and two-dimensional gel electrophoresisELECTROPHORESIS, Issue 16 2003Gianfranco Mamone Abstract We report a fast and easy-to-use procedure that combines polyacrylamide gel electrophoresis with matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF) and nanoelectrospray-tandem mass spectrometry (nES-MS/MS) analysis for the identification of casein components and defined phosphorylated sites. This methodology ensured identification of more than 30 phosphorylated proteins, five ,-, fifteen ,s1 -, ten ,s2 -, and four ,-casein (CN) components, including nonallelic, differently phosphorylated, and glycosylated forms. The sugar motif covalently bound to ,-CN was identified as chains, trisaccharide GalNAc, Gal, NeuGc, and tetrasaccharide 1GalNAc, 1Gal, 2NeuGc. Also identified was a biantennary chain made up of both chains of trisaccharide 1GalNAc, 1Gal, 1NeuGc, and tetrasaccharide 1GalNAc, 1Gal, 2NeuGc moiety on a single ,-CN component. The phosphate group on site Ser12 of tryptic peptide 8,22 of most phosphorylated ,s1 -CN (11 phosphate groups) was localized and the oligosaccharide sequence of the main tryptic glycopeptides of two ,-CN components was determined by means of MS/MS analysis. [source] Differential alteration of lipid antigen presentation to NKT cells due to imbalances in lipid metabolismEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 6 2007Jens Schümann Abstract Deficiencies in enzymes of the lysosomal glycosphingolipid degradation pathway or in lysosomal lipid transfer proteins cause an imbalance in lipid metabolism and induce accumulation of certain lipids. A possible impact of such an imbalance on the presentation of lipid antigens to lipid-reactive T cells has only been hypothesized but not extensively studied so far. Here we demonstrate that presentation of lipid antigens to, and development of, lipid-reactive CD1d-restricted NKT cells, are impaired in mice deficient in the lysosomal enzyme ,-galactosidase (,Gal) or the lysosomal lipid transfer protein Niemann-Pick C (NPC) 2. Importantly, the residual populations of NKT cells selected in ,Gal,/, and NPC2,/, mice showed differential TCR and CD4 repertoire characteristics, suggesting that differential selecting CD1d:lipid antigen complexes are formed. Furthermore, we provide direct evidence that accumulation of lipids impairs lipid antigen presentation in both cases. However, the mechanisms by which imbalanced lipid metabolism affected lipid antigen presentation were different. Based on these results, the impact of lipid accumulation should be generally considered in the interpretation of immunological deficiencies found in mice suffering from lipid metabolic disorders. [source] RNA interference reveals a role for TLR2 and TLR3 in the recognition of Leishmania donovani promastigotes by interferon,,-primed macrophagesEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 2 2006Jean-Frédéric Flandin Abstract Leishmania donovani promastigotes evade the induction of a proinflammatory response during their invasion of naive macrophages. However, their entry into IFN-,-primed macrophages is accompanied by the secretion of nitric oxide (NO) and proinflammatory cytokines. In the present study, we addressed the hypothesis that priming with IFN-, induces the expression of a receptor that enables mouse macrophages to recognize L. donovani promastigotes. We observed that in IFN-,-primed macrophages, L. donovani promastigotes stimulated Interleukin-1 receptor-associated kinase-1 (IRAK-1) activity. We next showed that Toll-like receptor (TLR)3 is barely detectable in naive macrophages but is expressed in IFN-,-treated macrophages. Silencing of TLR3, TLR2, IRAK-1 and myeloid differentiation factor 88 (MyD88) expression by RNA interference revealed that both TLR are involved in the secretion of NO and TNF-, induced by L. donovani promastigotes. Using L. donovani mutants, we showed that TLR2-mediated responses are dependent on Gal,1,4Man,-PO4 -containing phosphoglycans, whereas TLR3-mediated responses are independent of these glycoconjugates. Furthermore, our data indicate a participation of TLR2 and TLR3 in the phagocytosis of L. donovani promastigotes and a role for TLR3 in the leishmanicidal activity of the IFN-,-primed macrophages. Collectively, our data are consistent with a model where recognition of L. donovani promastigotes depends on the macrophage activation status and requires the expression of TLR3. [source] Intracellular trafficking and release of intact edible mushroom lectin from HT29 human colon cancer cellsFEBS JOURNAL, Issue 7 2000Lu-Gang Yu Our previous studies have shown that the Gal,1,3GalNAc,- (Thomsen,Friedenreich antigen)-binding lectin from the common edible mushroom Agaricus bisporus (ABL) reversibly inhibits cell proliferation, and this effect is a consequence of inhibition of nuclear localization sequence-dependent nuclear protein import after ABL internalization [Yu, L.G., Fernig, D.G., White, M.R.H., Spiller, D.G., Appleton, P., Evans, R.C., Grierson, I., Smith, J.A., Davies, H., Gerasimenko, O.V., Petersen, O.H., Milton, J.D. & Rhodes, J.M. (1999) J. Biol. Chem.274, 4890,4899]. Here, we have investigated further the intracellular trafficking and fate of ABL after internalization in HT29 human colon cancer cells. Internalization of 125I-ABL occurred within 30 min of the lectin being bound to the cell surface. Subcellular fractionation after pulse labelling of the cells with 125I-ABL for 2 h at 4 °C followed by culture of the cells at 37 °C demonstrated a steady increase in radioactivity in a crude nuclear extract. The radioactivity in this extract reached a maximum after 10 h and declined after 20 h. Release of ABL from the cell, after pulse labelling, was assessed using both fluorescein isothiocyanate-labelled ABL and 125I-ABL and was slow, with a t1/2 of 48 h. Most of the 125I-ABL both inside cells and in the medium remained intact, as determined by trichloroacetic acid precipitation and SDS/PAGE, and after 48 h only 22 ± 2% of ABL in the medium and 14 ± 2% inside the cells was degraded. This study suggests that the reversibility of the antiproliferative effect of ABL is associated with its release from cells after internalization. The internalization and subsequent slow release, with little degradation of ABL, reflects the tendency of lectins to resist biodegradation and implies that other endogenous or exogenous lectins may be processed in this way by intestinal epithelial cells. [source] A New Lignan and Four New Lignan Glycosides from Mananthes patentifloraHELVETICA CHIMICA ACTA, Issue 2 2006Junmian Tian Abstract A new lignan, 5-hydroxyjusticidin,A (=,9-(1,3-benzodioxol-5-yl)-5-hydroxy-4,6,7-trimethoxynaphtho[2,3- c]furan-1(3H)-one; 1), and four new diphyllin-type lignan glycosides, mananthosides C,F (2,5), containing glucosyl (Glc), arabinosyl (Ara), galactosyl (Gal), and/or apiosyl residues, have been isolated from Mananthes patentiflora, together with five known compounds. Their structures and configurations were elucidated by in-depth 1D- and 2D-NMR experiments, as well as MS analysis. [source] Glycosylation status of haptoglobin in sera of patients with prostate cancer vs. benign prostate disease or normal subjectsINTERNATIONAL JOURNAL OF CANCER, Issue 1 2008Tsutomu Fujimura Abstract We studied chemical level and glycosylation status of haptoglobin in sera of patients with prostate cancer, as compared to benign prostate disease and normal subjects, with the following results. (i) Haptoglobin level was enhanced significantly in sera of prostate cancer. (ii) Sialylated bi-antennary glycans were the dominant structures in haptoglobins from all 3 sources, regardless of different site of N-linked glycan. The N-linked glycans at N184 were exclusively bi-antennary, and showed no difference between prostate cancer vs. benign prostate disease. (iii) Tri-antennary, N-linked, fucosylated glycans, carrying at least 1 sialyl-Lewisx/a antenna, were predominantly located on N207 or N211 within the amino acid 203-215 sequence of the ,-chain of prostate cancer, and were minimal in benign prostate disease. Fucosylated glycans were not observed in normal subjects. A minor tri-antennary N-linked glycan was observed at N241 of the ,-chain in prostate cancer, which was absent in benign prostate disease. (iv) None of these N-linked structures showed the expected presence of disialylated antennae with GalNAc,4(NeuAc,3)Gal,3(NeuAc,6)GlcNAc,Gal, or its analogue, despite cross-reactivity of prostate cancer haptoglobin with monoclonal antibody RM2. (v) Minor levels of O -glycosylation were identified in prostate cancer haptoglobin for the first time. Mono- and disialyl core Type 1 O-linked structures were identified after reductive ,-elimination followed by methylation and mass spectrometric analysis. No evidence was found for the presence of specific RM2 or other tumor-associated glycosyl epitopes linked to this O -glycan core. In summary, levels of haptoglobin are enhanced in sera of prostate cancer patients, and the N -glycans attached to a defined peptide region of its ,-chain are characterized by enhanced branching as well as antenna fucosylation. © 2007 Wiley-Liss, Inc. [source] X-ray structural analysis of the ligand-recognition mechanism in the dual-affinity labeling of c-type lysozyme with 2,,3,-epoxypropyl ,-glycoside of N -acetyllactosamineJOURNAL OF MOLECULAR RECOGNITION, Issue 2 2003Michiro Muraki Abstract In spite of the belonging to the same c-type lysozyme family, hen egg-white lysozyme (HEWL) was much less susceptible to the dual-affinity labeling with 2,,3,-epoxypropyl ,-glycoside of N -acetyllactosamine (Gal,1,4GlcNAc-Epo) than human lysozyme (HL). The three-dimensional structures of the HEWL labeled with single Gal,1,4GlcNAc-Epo and the Glu102-mutant HL labeled with double Gal,1,4GlcNAc-Epo were determined by X-ray crystallography at resolutions of 1.85 and 2.0,Å, respectively. The overall conformation and the interaction mode of the carbohydrate ligand part in the singly labeled HEWL and the doubly labeled Glu102-mutant HL were basically identical to those of the correspondingly labeled wild-type HL with minor alterations in some stereochemical parameters. A detailed comparison of the structures revealed the key protein,carbohydrate and carbohydrate,carbohydrate interactions essential for the dual labeling. It was suggested that the difference in the efficiency of the dual labeling was caused by the structural difference between Gln104 in HL and Asn103 in HEWL. The relevance to our previous study and the carbohydrate,carbohydrate interaction on cell-surface membranes were discussed. Copyright © 2003 John Wiley & Sons, Ltd. [source] Use of perfluorocarbon (fluorinert) to enhance reporter gene expression following intratracheal instillation into the lungs of Balb/c mice: Implications for nebulized delivery of plasmidsJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 9 2001Aditya Das Abstract Perfluorocarbons combine high respiratory gas dissolving capabilities with extreme chemical and biological inertness and therefore offer an attractive option as an excipient in the area of pulmonary therapeutics. Perfluorocarbons have also been shown to ,float' mucus, because of their high densities (1.9,2.5 g/mL), which may hold potential in gene delivery for cystic fibrosis patients, in terms of enhancing penetration through highly viscous mucus and thereby providing access to target epithelial cells to correct the gene defect. Additionally, their low surface tension allows for better dispersion. A commonly available perflurocarbon, heptacosafluorotributylamine (Fluorinert), was used to deliver either plasmid DNA (pDNA) alone or cationic-lipid-complexed plasmid DNA to the lungs of Balb/c mice by direct intratracheal instillation. The complexes consisted of supercoiled (SC) plasmid DNA (4.7 Kb, 0.625 mg/mL) and lipid (ethyldimyristoyl phosphatidylcholine [EDMPC]/cholesterol [1:1 mole ratio], with pDNA (3:1 mg pDNA/mM EDMPC in 20 mM Tris-HCl pH 8.0) expressing chloramphenicol acetyl transferase (CAT) or ,-galactosidase (,-Gal). pDNA alone was supplemented with 14% w/v Fluorinert. Cationic lipid/pDNA complexes were supplemented with 3, 8, and 14% w/v Fluorinert. Results showed that the CAT expression from pDNA alone was enhanced 24,× using 14% w/v Fluorinert, whereas that from the cationic-lipid-formulated pDNA was enhanced 7,× using 14% w/v Fluorinert. Immunohistochemistry showed that ,-Gal expression was primarily from epithelial cells and not from F4/80 or MAC3 antigen-stained cells (predominantly macrophages), indicating efficient delivery. © 2001 Wiley-Liss, Inc. and the American Pharmaceutical Association J Pharm Sci 90:1336,1344, 2001 [source] Biostability and pharmacokinetics of LJP 920, an octameric Gal (,1,3) Gal conjugate for the inhibition of xenotransplantation rejectionJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 7 2001Lee Jia Antibodies to an ,-galactosyl saccharide structure present in human serum are associated with hyperacute rejection and delayed xenograft rejection after pig-to-primate xenotransplantation. To overcome this major barrier to the xenotransplantation, LJP 920, a galactosyl ,1,3 galactose (Gal (,1,3) Gal) coupled to a non-immunogenic platform at a valency of eight Gal (,1,3) Gal molecules/platform, was synthesized to clear circulating antibodies and to inhibit their production by B cells that produce these antibodies. Herein we report on the stability of LJP 920 in biological media and its pharmacokinetic profile. Incubation of LJP 920 with mouse serum or liver microsomes at 37°C for 2 days showed no indication of degradation of the conjugate as detected by a reversed-phase HPLC method, indicating that the conjugate is not subject to enzymatic metabolism. After intravenous administration of LJP 920 to mice at the doses of 20 and 100 mg kg,1, LJP 920 serum concentration decreased rapidly, showing a biphasic pattern, with a distribution half-life of 3 min and an elimination half-life of more than 30 min, respectively. The serum-to-erythrocyte concentration ratio of LJP 920 was 33- and 36-fold excess at 0.5 and 5 min, respectively, after intravenous administration (100 mg kg,1). Both Cmax and AUC values increased in a dose-proportional manner. LJP 920 displayed a great distribution to well-perfused tissues. It was eliminated mainly through renal excretion in the unchanged form, which accounted for 23% of the total amount within 8 h of dosing. [source] Atorvastatin or transgenic expression of TFPI inhibits coagulation initiated by anti-nonGal IgG binding to porcine aortic endothelial cellsJOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 9 2010C. C. LIN Summary.,Background:,Intravascular thrombosis remains a barrier to successful xenotransplantation. Tissue factor (TF) expression on porcine aortic endothelial cells (PAECs), which results from their activation by xenoreactive antibodies (Abs) to Gal,1,3Gal (Gal) and subsequent complement activation, plays an important role. Objectives:,The present study aimed to clarify the role of Abs directed against nonGal antigens in the activation of PAECs to express functional TF and to investigate selected methods of inhibiting TF activity. Methods:,PAECs from wild-type (WT), ,1,3-galactosyltransferase gene-knockout (GT-KO) pigs, or pigs transgenic for CD46 or tissue factor pathway inhibitor (TFPI), were incubated with naïve baboon serum (BS) or sensitized BS (with high anti-nonGal Ab levels). TF activity of PAECs was assessed. Results:,Only fresh, but not heat-inactivated (HI), naïve BS activated WT PAECs to express functional TF. Similarly, PAECs from CD46 pigs were resistant to activation by naïve BS, but not to activation by fresh or HI sensitized BS. HI sensitized BS also activated GT-KO PAECs to induce TF activity. TF expression on PAECs induced by anti-nonGal Abs was inhibited if serum was pretreated with (i) an anti-IgG Fab Ab or (ii) atorvastatin, or (iii) when PAECs were transgenic for TFPI. Conclusions:,Anti-nonGal IgG Abs activated PAECs to induce TF activity through a complement-independent pathway. This implies that GT-KO pigs expressing a complement-regulatory protein may be insufficient to prevent the activation of PAECs. Genetic modification with an ,anticoagulant' gene (e.g. TFPI) or a therapeutic approach (e.g. atorvastatin) will be required to prevent coagulation dysregulation after pig-to-primate organ transplantation. [source] Impaired sexual behavior in male mice deficient for the ,1,3 N -Acetylglucosaminyltransferase-I geneMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 5 2008Franziska Biellmann Abstract The ,1,3 N -acetylglucosaminyltransferase-1 (B3gnt1) gene encodes a poly- N -acetyllactosamine synthase which can initiate and extend poly- N -acetyllactosamine chains [Gal(,1,4)GlcNAc (,1,3)n]. Previous investigations with heterozygous and homozygous null mice for this gene have revealed the importance of poly- N -acetyllactosamine chains for the formation of olfactory axon connections with the olfactory bulb and the migration of gonadotropin releasing hormone neurons to the hypothalamus. The possible long-term effects of these developmental defects, however, has not yet been studied. Here we have examined a reproductive phenotype observed in B3gnt1 -null mice. Whereas the B3gnt1 null females were fertile, the B3gnt1 null males were not able to sire litters at the expected rate when mated to either wildtype or B3gnt1 -null females. We assessed male sexual behavior as well as male reproduction parameters such as testes size, spermatogenesis, sperm number, morphology, and the development of early embryos in order to identify the source of a reduced rate of reproduction. Our findings show that the B3gnt1 null male reproductive organs were functional and could not account for the lower rate at which they produced offspring with wildtype conspecifics. Hence, we propose that the phenotype observed resulted from an impaired sexual response to female mating partners. Mol. Reprod. Dev. 75: 699,706, 2008. © 2007 Wiley-Liss, Inc. [source] Role of sialic acid in bovine sperm,zona pellucida bindingMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 5 2007José Guillermo Velásquez Abstract Sperm binding activity has been detected in zona pellucida (ZP) glycoproteins and it is generally accepted that this activity resides in the carbohydrate moieties. In the present study we aim to identify some of the specific carbohydrate molecules involved in the bovine sperm,ZP interaction. We performed sperm binding competition assays, in vitro fecundation (IVF) in combination with different lectins, antibodies and neuraminidase digestion, and chemical and cytochemical analysis of the bovine ZP. Both MAA lectin recognising ,-2,3-linked sialic acid and neuraminidase from Salmonella typhimurium with catalytic activity for ,-2,3-linked sialic acid, demonstrated a high inhibitory effect on the sperm,ZP binding and oocyte penetration. These results suggest that bovine sperm,ZP binding is mediated by ,-2,3-linked sialic acid. Experiments with trisaccharides (sialyllactose, 3,-sialyllactosamine and 6,-sialyllactosamine) and glycoproteins (fetuin and asialofetuin) corroborated this and suggest that at least the sequence Neu5Ac(,2-3)Gal(,1-4)GlcNAc is involved in the sperm,ZP interaction. Moreover, these results indicate the presence of a sperm plasma membrane specific protein for the sialic acid. Chemical analysis revealed that bovine ZP glycoproteins contain mainly Neu5Ac (84.5%) and Neu5GC (15.5%). These two types of sialic acid residues are probably linked to Gal,1,4GlcNAc and GalNAc by ,-2,3- and ,-2,6-linkages, respectively, as demonstrated by lectin cytochemical analysis. The use of a neuraminidase inhibitor resulted in an increased number of spermatozoa bound to the ZP and penetrating the oocyte. From this last result we hypothesize that a neuraminidase from cortical granules would probably participate in the block to polyspermy by removing sialic acid from the ZP. Mol. Reprod. Dev. 74: 617,628, 2007. © 2006 Wiley-Liss, Inc. [source] Privileging masculinity in the social construction of Basque identityNATIONS AND NATIONALISM, Issue 3 2001Begoña Echeverria Following a framework developed by Susan Gal and Judith Irvine (1995), this article illustrates how Basque-medium schools promulgate an androcentric vision of the Basque nation. First, male privilege is upheld in textbooks through the erasure of women's contributions to Basque language and culture, so that men appear as the quintessential Basque speakers and cultural agents. Secondly, language ideologies about Spanish and Basque recursively construct Basque ethnic identity is such a way that it centres on vernacular Basque, whose primary marker is a second person pronoun, ,hi', which indirectly indexes male speakers and masculinity. An iconic relationship is thereby created between authentic Basque identity, Basque culture, Basque linguistic forms and masculinity. However, I also show that women have challenged this male privilege in various domains, thereby opening up the possibility of a Basque nation that embraces its female as well as its male members. [source] "Black Gal Swing": Color, Class, and Category in Globalized CultureAMERICAN ANTHROPOLOGIST, Issue 1 2001Fred J. Hay Scattered Belongings: Cultural Paradoxes of "Race," Nation and Gender. Jayne O. Ifekwunigwe. New York: Routledge, 1999. 221 pp. Race and Ideology: Language, Symbolism, and Popular Culture. Arthur K. Spears. ed. Detroit, MI: Wayne State University Press, 1999. 242 pp. Spurious Issues: Race and Multiracial Identity Politics in the United States. Rainier Spencer. Boulder, CO: Westview Press, 1999. 222 pp. [source] Differential expression of glycans in the hippocampus of rats trained on an inhibitory learning paradigmNEUROPATHOLOGY, Issue 6 2006Alejandra Hidalgo The glycan chains of glycoconjugates play important roles in cell,cell and cell,matrix interactions. In the CNS, previous studies on learning and memory suggest the importance of oligosaccharides attached to glycoconjugates in the modulation of synaptic connections. We studied the hippocampal glycan distribution of rats subject to an inhibitory avoidance task. The expression of glycans was examined by lectin-histochemistry using Vicia villosa lectin (VVL) for terminal ,/, N-acetylgalactosamine (,/, GalNAc); Galanthus nivalus lectin (GNL) for terminal mannose ,-1,3 (Man ,-1,3); Peanut agglutinin (PNA) for galactose ,-1,3N-acetylgalactosamine (Gal ,-1,3 GalNAc); Erythrina cristagalli lectin (ECL) for galactose ,-1,4 N-acetylglucosamine (Gal ,-1,4 GlcNAc); Sambucus nigra lectin (SNA) for sialic acid ,-2.6 galactose (SA ,-2,6 Gal); Maackia amurensis lectin II (MAL II) for sialic acid ,-2,3 (SA ,-2,3); Wheat germ agglutinin (WGA) for terminal N-acetylglucosamine with/without sialic acid (GlcNAc wo SA); succynilated WGA (sWGA) for terminal N-acetylglucosamine without sialic acid (terminal GlcNAc without SA); Griffonia simplicifolia lectin II (GSL II) for terminal ,/, N-acetylglucosamine (,/, GlcNAc terminal); and Lotus tetragonolobus lectin (LTL) ,,fucose. Two groups of 10 animals were examined: non-trained (Control) and Trained rats. ECL, sWGA and GSL II were negative for both groups in all the hippocampal subfields studied. For both groups, VVL was negative in CA4 and granular cells of the Dentate Gyrus (DG) and LTL was negative in the CA4 subfield. Expression of ,/, GalNAc, , -fucose and GlcNAc in other hippocampal subfields was positive, with no differences between groups. However, expression of Man ,-1,3 was significantly higher in the CA1, CA2, CA3, and CA4 subfields in the Trained group. On the other hand, expression of Gal ,-1,3 GalNAc was significantly low in CA4 and DG in the Trained group. In conclusion, the results here presented indicate that the exposure of rats to an associative behavioral paradigm related to declarative memory, involves some regulatory mechanism/s for the differential patterns of glycan expression. [source] Insecticidal action of mammalian galectin-1 against diamondback moth (Plutella xylostella)PEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 8 2009Shiang Jiuun Chen Abstract BACKGROUND: Previous studies showed that mammalian galectin-1 (GAL1) could interact with chitosan or chitin, one component of the peritrophic membrane (PM). This finding suggests that the PM could be a target of GAL1, which prompted the authors to explore the effect of GAL1 on larval growth and its potential mechanism. RESULTS: The development of Plutella xylostella (L.) larvae was significantly disturbed after they were fed recombinant GAL1. The histochemical structure and immunostaining pattern suggested that GAL1 treatment resulted in dose- and time-dependent disruption of the microvilli and abnormalities in these epithelial cells. Ultrastructural studies showed that the PM was not present in the midgut of GAL1-treated insects; instead, numerous bacteria were found in the lumen area. These results indicate that the protective function of the PM was disrupted by GAL1 treatment. Moreover, in vitro data showed that GAL1 interacts with chitosan/chitin in a dose-dependent manner, and also specifically binds to the PM in vitro. CONCLUSION: In view of the fact that the carbohydrate recognition domain of GAL1 recognises the structural motif N -acetyl lactosamine (Gal , 1,4 GlcNAc), which is similar to that of chitin (,-1,4 N -acetyl- D -glucosamine), it is proposed that the insecticidal mechanism of GAL1 involves direct binding with chitin to interfere with the structure of the PM. Copyright © 2009 Society of Chemical Industry [source] Energetics of galactose, and glucose,aromatic amino acid interactions: Implications for binding in galactose-specific proteinsPROTEIN SCIENCE, Issue 9 2004Mannargudi S. Sujatha Abstract An aromatic amino acid is present in the binding site of a number of sugar binding proteins. The interaction of the saccharide with the aromatic residue is determined by their relative position as well as orientation. The position-orientation of the saccharide relative to the aromatic residue was found to vary in different sugar-binding proteins. In the present study, interaction energies of the complexes of galactose (Gal) and of glucose (Glc) with aromatic residue analogs have been calculated by ab initio density functional (U-B3LYP/ 6-31G**) theory. The position-orientations of the saccharide with respect to the aromatic residue observed in various Gal-, Glc-, and mannose,protein complexes were chosen for the interaction energy calculations. The results of these calculations show that galactose can interact with the aromatic residue with similar interaction energies in a number of position-orientations. The interaction energy of Gal,aromatic residue analog complex in position-orientations observed for the bound saccharide in Glc/Man,protein complexes is comparable to the Glc,aromatic residue analog complex in the same position-orientation. In contrast, there is a large variation in interaction energies of complexes of Glc- and of Gal- with the aromatic residue analog in position-orientations observed in Gal,protein complexes. Furthermore, the conformation wherein the O6 atom is away from the aromatic residue is preferred for the exocyclic ,CH2OH group in Gal,aromatic residue analog complexes. The implications of these results for saccharide binding in Gal-specific proteins and the possible role of the aromatic amino acid to ensure proper positioning and orientation of galactose in the binding site have been discussed. [source] | |||||||||||||||||||||||||||||||||||||