Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.2.1.23 (beta-galactosidase)
14,648 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The distribution and localization of acid stable trypsin inhibitor (ASTI) in normal and malignant human tissues from various organs were examined using immunohistochemical techniques that used goat antibody raised against highly purified ASTI from human urine. Tissues were assessed as positive only when they were stained by both the biotin-avidin-peroxidase complex system and biotin-streptavidin-beta-galactosidase complex system, and the staining was abolished by absorption with purified ASTI. Under normal conditions, ASTI immunoreactivity was observed in only a few organs. Positive tissues for ASTI immunoreactivity included the kidney proximal tubules, glial cells of the cerebrum, fibrillar structures of the lamina propria of the stomach and colon, and bronchial epithelial cells. No ASTI immunoreactivity was observed in the cardiovascular system, reproductive system, or other tissues examined. As is not the case for normal tissues, ASTI immunoreactivity was found to be widely distributed in malignant tumors. Staining was observed in the extracellular space, i.e., in the stroma of the tumor and in connective tissues around the tumor invasion, whereas no ASTI immunoreactivity was detected in the malignant cells. Considering the identity of the first 36 NH2-terminal residues of ASTI purified from plasma or urine with a recently reported endothelial cell growth factor, the present findings suggest that ASTI could play an important role, not limited to its function as a protease inhibitor, in the invasive growth of malignant neoplasms.
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PMID:Distribution of acid stable trypsin inhibitor immunoreactivity in normal and malignant human tissues. 247 69

The relationships both between cholinergic neurons and substance P (SP) and between cholinergic neurons and calcitonin gene-related peptide (CGRP) terminals were examined in the rat sacral intermediolateral nucleus at the light and electron microscopic levels by means of double-immunostaining methods. Cholinergic neurons were labeled by a monoclonal antibody to choline acetyltransferase (CAT) with the avidin-biotin technique and stained bluish-green by indolyl-beta-galactoside reaction products with beta-galactosidase as a marker. On the same sections, SP or CGRP fibers were labeled by polyclonal antisera to SP or CGRP after application of the peroxidase-antiperoxidase (PAP) method and stained brown by the p-dimethylaminoazobenzene (DAB) reaction. After embedding in Epon, light and electron microscopic sections were examined. At the light microscopic level, CGRP-like immunoreactive (CGRP-I) fibers and SP-like immunoreactive (SP-I) fibers were found to pass through the lateral edge of the dorsal horn and then into the dorsal region of the sacral intermediolateral nucleus. In addition, SP-I fibers also extend from the dorsolateral funiculus into the entire sacral intermediolateral region. At the electron microscopic level, many axosomatic and axodendritic synapses were found between CAT-I structures and SP-I terminals in the intermediolateral nucleus, whereas most of the CGRP-I terminals in this area made axodendritic synapses with CAT-I dendrites. These results indicate that cholinergic neurons in the sacral intermediolateral nucleus receive direct synaptic input from SP-I and CGRP-I terminals.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Interaction between cholinergic neurons and substance P or calcitonin gene-related peptide terminals of the rat sacral intermediolateral nucleus: double immunostaining at the light and electron microscopic levels. 247 8

Using a combination of horseradish peroxidase (HRP) histochemistry and immuno-beta-galactosidase staining, and pre-embedding immuno-electron microscopy, the present study was intended to re-examine the origins of enkephalin-like immunoreactivity (ENKI) in the rat lateral septum (LS), and to show the fine structures of septal ENKI fibers. Following HRP injection into the LS, double-labeled cells which contained a homogeneous blue reaction product of ENKI and a black or brown granular reaction product of retrogradely transported HRP were identified in 4 discrete brain regions: perifornical hypothalamic area at the level of the paraventricular nucleus (PeF); posterior part of the anterior hypothalamic nucleus (AHP); bed nucleus of the stria terminalis, medial division, posterolateral part (BSTMPL); and dorsal hypothalamic area (DA). Immuno-electron micrographs demonstrated that some of the ENKI terminals in the LS form synapses with the soma and dendrites of septal neurons devoid of ENKI, though ENKI dendrites postsynaptic to non-immunoreactive terminals were also seen in the LS. These findings suggest that a large proportion of septal ENKI fibers have their origins in the above regions (PeF, AHP, BSTMPL, DA) different from the previously considered one, and they further provide a morphological basis for the postsynaptic inhibitory effects of enkephalins on septal neurons.
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PMID:Reassessment of enkephalin (ENK)-containing afferents to the rat lateral septum with reference to the fine structures of septal ENK fibers. 249 65

Assays for alkaline phospatase, beta-galactosidase, penicillinase and peroxidase were optimised for quantitation in microtitre plate wells. Their value as labels in microtitre plate enzymeimmunoassay (EIA) for progesterone was assessed following coupling with 11 alpha-hydroxyprogesterone 11-glucuronide using an active ester procedure. Bridge-heterologous antiserum (11 alpha-hydroxyprogesterone 11-hemisuccinate-bovine serum albumin as immunogen) was used to minimize bridge recognition. The limits of detection of the enzymes were in the order penicillinase greater than peroxidase greater than alkaline phosphatase greater than beta-galactosidase. Under appropriate conditions it was possible to achieve greater than 50% displacement of label with 50 pg of progesterone for all four labels.
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PMID:A comparison of alkaline phosphatase, beta-galactosidase, penicillinase and peroxidase used as labels for progesterone determination in milk by heterologous microtitre plate enzymeimmunoassay. 250 94

Seven antisera raised against 11 alpha-hydroxyprogesterone 11-hemisuccinate (P11-HS) were used in microtitre plate enzymeimmunoassays (EIAs) for progesterone to identify improvements in sensitivity achievable by using various heterologous labels. EIAs using beta-galactosidase linked to P11-HS, 11 alpha-hydroxyprogesterone 11-hemimaleate (P11-HM), 11 alpha-hydroxyprogesterone 11-glucuronide (P11-Glu) or progesterone 3-(o-carboxymethyl) oxime (P3-CMO) were compared. Loss of sensitivity through bridge recognition was least evident using the P11-Glu derivative. The same seven antisera were used to evaluate assay sensitivity using beta-galactosidase, alkaline phosphatase, penicillinase and peroxidase linked to P11-HS or P11-Glu as label. Consistent improvements were achieved with the heterologous assays in the order penicillinase greater than alkaline phosphatase/peroxidase greater than beta-galactosidase: with penicillinase, sensitivity generally exceeded that of RIA. These data provide evidence for the general efficacy of the combination 11 alpha-hemisuccinate (immunogen bridge) and 11 alpha-glucuronide (label bridge) in reducing bridge recognition. EIA performed at 4 degrees C provided greater sensitivity than at ambient temperature (21 degrees C) or 40 degrees C, however, ambient temperature incubation provided a practical compromise. Equilibrium was not achieved under any of the conditions investigated.
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PMID:The influence of heterology, enzyme label and assay conditions on the sensitivity of microtitre plate enzymeimmunoassays for progesterone in milk. 255 Jul 5

Relationships between leucine-enkephalin fibers and cholinergic neurons in the rat sacral intermediolateral nucleus were examined by light and electron microscopy using double-immunostaining method. Cholinergic neurons in the sacral intermediolateral nucleus were labeled by a rat-mouse monoclonal antibody to choline acetyltransferase and stained bluish green with 5-bromo-4-chloro-3-indolyl-beta-D- galactoside reaction products using beta-galactosidase as a marker. On the same sections, leucine-enkephalin fibers were labeled by a rabbit polyclonal antiserum to leucine-enkephalin and stained brown by diaminobenzidine reaction products using peroxidase as a marker. After embedding in Epon, the sections were examined in light and electron microscopes. In the light microscope, choline acetyltransferase-like immunoreactive cells were seen in the sacral intermediolateral nucleus. In the same region, leucine-enkephalin-like immunoreactive cells. In the electron microscope, 5-bromo-4-chloro-3-indolyl-beta-D-galactoside reaction products were in the form of coarse electron dense deposits in the choline acetyltransferase-like immunoreactive structures and could be distinguished from the much finer grained diaminobenzidine reaction products. Choline acetyltransferase-like immunoreactive neurons received synaptic inputs from leucine-enkephalin fibers-like immunoreactive terminals. These findings suggest that leucine-enkephalin fibers may affect the activity of cholinergic parasympathetic preganglionic neurons.
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PMID:Enkephalin fibers synapse on cholinergic neurons in the rat sacral intermediolateral nucleus: a double-immunostaining at the light and electron microscopic levels. 264 54

The sensitivities of monoclonal antibody-based enzyme immunoassays for 11-deoxycortisol using alkaline phosphatase (AP), horseradish peroxidase (HRP), beta-galactosidase (beta-GAL) and glucose oxidase (GOD) as labels were compared. The anti-11-deoxycortisol antibody used was that produced in ascites by inoculating antibody-secreting hybridoma cells into mice. Enzyme labeling of 11-deoxycortisol was carried out by the N-succinimidyl ester method. The activated ester of 4-(2-carboxyethylthio)-11-deoxycortisol was treated with each enzyme to give a homologous enzyme-labeled antigen. In the competitive immunoassay, the bound and free enzyme-labeled antigens were separated by a double antibody method and the enzymic activity of the immune precipitate was determined by colorimetric and fluorimetric methods. The AP activity was measured in three ways, using p-nitrophenyl phosphate, nicotinamide adenine dinucleotide phosphate (NADP), and 4-methylumbelliferyl phosphate as substrates. o-Nitrophenyl beta-D-galactopyranoside and 4-methylumbelliferyl beta-D-galactopyranoside were used for beta-GAL, and 3,3',5,5'-tetramethylbenzidine (TMB) and 3-(p-hydroxyphenyl)propionic acid (HPPA) for HRP. In the case of GOD, TMB and HPPA were used in combination with HRP. A dose-response curve with a high sensitivity was obtained in each 11-deoxycortisol assay system by the use of a minimum amount of the enzyme-labeled antigen at an appropriate dilution of monoclonal anti-11-deoxycortisol antibody (Ka = 2 x 10(10) M-1). The amounts of 11-deoxycortisol needed to displace 50% of the bound label ranged from 5 to 15 pg in the colorimetric methods, and 4-9 pg in the fluorimetric methods.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Sensitivity of steroid enzyme immunoassays. Comparison of four label enzymes in an assay system using a monoclonal anti-steroid antibody. 268 Jan 24

The hybridoma, 62H3, which secretes a monoclonal IgG2b with anti-HLA-DR specificity, was expanded in pristane-primed BALB/c mice and the antibody was isolated from the ascitic fluid by affinity chromatography on Protein A-Sepharose. The purified IgG2b antibody was tested by an enzyme immunoassay for antibody activity against a panel of 40 self and non-self antigens. It was found to react strongly with beta-galactosidase, actin, glutamate dehydrogenase, rabbit and human IgG and di- and trinitrophenyl groups; and moderately with tubulin, insulin and phosphorylcholine; but it did not react with various other self and non-self antigens, such as DNA, albumin, keyhole limpet hemocyanin, hen lysozyme and horseradish peroxidase. Fab and Fc fragments were prepared from this IgG2b by papain proteolysis. The Fab fragment possessed the same spectrum of polyreactivities as the native IgG2b, whereas no activity was detected with the Fc fraction. In order to investigate the properties of the antigen binding site, the actin, TNP and rabbit IgG antibody activities were studied in more detail by enzyme immunoassay, Western blot and immunocytochemistry. The monomolecular nature of this multireactivity was confirmed by immunoabsorption analysis. Furthermore, 62H3 monoclonality was also verified by comparative isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis with other monospecific antibodies. The dissociation constants (Kd) of antigen-antibody equilibria in solution were measured. The Kd for actin was 1.11 +/- 0.24 x 10(-5) M and the Kd for TNP-BSA was 8.7 +/- 0.51 x 10(-7) M. No interaction with rabbit IgG could be detected in solution. These findings raise the question of the possible implication in autoimmune pathology or in normal physiology of IgG class polyspecific antibodies with solid-phase restricted cross-reactive rheumatoid factor activity.
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PMID:Immunochemical studies of a murine polyreactive IgG2b autoantibody with rheumatoid factor activity. 277 Jul 48

The co-localization of corticotropin-releasing factor (CRF)- and enkephalin (ENK)-like immunoreactivities in nerve cells of the rat hypothalamus and adjacent areas was investigated by the simultaneous application of immuno-beta-galactosidase staining and the peroxidase-antiperoxidase (PAP) method to the same sections. CRF-like immunoreactive cells were stained blue with immuno-beta-galactosidase staining and ENK-like immunoreactive cells brown with the PAP method. Double-labeled cells with overlap of blue and brown immunoreaction products were identified in all subregions of the paraventricular hypothalamic nucleus (PVH), in contrast to previous studies showing the occurrence of double-labeled cells only in the parvocellular part of the PVH. Other areas that contained double-labeled cells were: the medial preoptic area, bed nucleus of the stria terminalis, periventricular hypothalamic nucleus, lateral hypothalamic area, dorsal hypothalamic area and subincertal nucleus. These findings suggest that nerve cells with both CRF- and ENK-like immunoreactivities may be more actively involved in neuroendocrine regulation and neural transmission than previously considered.
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PMID:Co-localization of corticotropin-releasing factor- and enkephalin-like immunoreactivities in nerve cells of the rat hypothalamus and adjacent areas. 278 46

Retrovirus-mediated gene transfer was used to develop a method for introducing genes into primary cultures of adult rat hepatocytes. Subconfluent monolayers of hepatocytes, cultured in hormonally defined media on different matrix substrata, were infected with helper-free stocks of a replication-defective retrovirus that constitutively expresses high levels of beta-galactosidase. Retrovirus-mediated transduction was measured by two methods: (i) an in situ cytochemical stain that specifically detects the expression of viral expressed beta-galactosidase, and (ii) Southern blot analysis, which measures the relative copy number of integrated provirus. Maximal transduction efficiency of approximately equal to 25% was achieved when the cells were infected after 3 days in culture; matrix had little effect on transduction efficiency. Enzyme cytochemical (catalase and glucose 6-phosphatase) and peroxidase immunocytochemical (asialoglycoprotein and UDP-glucuronosyltransferase) analyses of the cultures indicated that greater than 95% of cells were hepatocytes. The demonstration of hepatocyte-specific organelles in cells expressing the viral-directed beta-galactosidase provided unambiguous evidence for the transduction of hepatocytes. These methods should be useful in the development of liver-directed somatic gene therapy and in the study of liver-specific gene regulation.
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PMID:Retrovirus-mediated transduction of adult hepatocytes. 283 28


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