Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.11.18 (MAP)
 7,412 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We describe the development of bi-specific monoclonal antibodies (MAb) capable of simultaneous recognition of rabbit immunoglobulin G (IgG) and horseradish peroxidase (HRP) for use in a variety of immunobased techniques. This bi-specific antibody, named McC8, was produced by fusion of the aminopterin-sensitive mouse hybridoma MAP.Ag.1, which secretes MAb against HRP, and splenocytes from a mouse previously immunized with whole rabbit IgG. The resultant hybrid-hybridoma co-dominantly expresses and secretes the immunoglobulin chains, i.e., IgG1 and IgG2b, of its respective parents, as determined by radial immunodiffusion. The binding sites on rabbit IgG for McC8 were determined on Western blots and in competition solid-phase enzymatic immunoassays with the use of allotype-specific rabbit sera. Both these techniques demonstrated that McC8 recognizes the light chain of the rabbit IgG molecule with preferential binding to the B4 kappa light-chain allotype. McC8 was successfully used in two-step immunocytochemistry for localization of calcitonin gene-related peptide (CGRP) in fibers of the superficial layers of the spinal trigeminal nucleus of the rat, as well as for localization of glial fibrillary acidic protein (GFAP)-immunoreactive sites in primary rat septal cell cultures, thus demonstrating its potential as a general developing reagent in conventional immunocytochemistry. McC8 compared favorably with peroxidase-antiperoxidase immunocytochemistry with respect to sensitivity. However, the bi-specific developing reagent proved superior to the conventional peroxidase-antiperoxidase procedure when both were employed in a similar fashion in tissues prone to display high background staining. Finally, McC8 was also employed as a developing reagent in a competitive ELISA designed for quantitation of CGRP with the use of a rabbit anti-CGRP primary antibody. The sensitivity of this quantitative ELISA (190 pg or 50 fmol CGRP per well) renders this bi-specific antibody suitable for use in quantitative immunoassays for detection of relevant peptides in biological systems.
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PMID:Development of a bi-specific monoclonal antibody for simultaneous detection of rabbit IgG and horseradish peroxidase: use as a general reagent in immunocytochemistry and enzyme-linked immunosorbent assay. 229 80

The injection of 25 mg/kg i.p. cyclosporin (CsA) for 3 wk caused marked functional and morphological deteriorations of pancreatic islet cells in Wistar rats that were prevented by the combined administration of p-aminobenzoic acid-N-D-mannoside sodium salt (K-MAP). In this article, the toxic effect of CsA on pancreatic islet cells and the preventive effect of K-MAP on CsA-associated islet cell toxicity were investigated. Prolonged hyperglycemia and depressed insulin secretion after the glucose challenge observed in CsA-treated rats could be prevented by the combined administration of 300 and 900 mg/kg K-MAP. Cytoplasmic vacuolizations and a decrease in the number of mitochondria, intact endoplasmic reticula, secretory granules, and insulin-positive cells, as revealed by peroxidase-antiperoxidase staining, could also be prevented by the administration of 900 mg/kg K-MAP. This preventive effect of K-MAP on CsA-associated islet cell toxicity may suggest the combined use of K-MAP with CsA in pancreas transplantation and treatment of insulin-dependent diabetes.
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PMID:Modulation of prostaglandin metabolism by K-MAP and prevention of toxic effect of cyclosporin on pancreatic islet cells. 264 33

Several procedures were employed to examine the in vitro interaction between S-100 proteins and microtubule proteins. Binding of S-100 to tau factors was observed under all experimental conditions. S-100 binding to microtubule-associated protein 2 (MAP2) was best detected by exposing nitrocellulose-immobilized MAP2 or MAPs to either 125I-labeled S-100 or biotinylated S-100. S-100 binding to tubulin was detected when the two protein fractions were first incubated with each other followed by exposure to the bifunctional cross-linker disuccinimidylsuberate, and then separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transfered onto nitrocellulose paper. By this procedure, complex formation between S-100 and tubulin, as well as between S-100 and a relatively low-molecular-weight MAP, was evidenced by immunoblotting using an anti-S-100 antiserum. Alternatively, complex formation between biotinylated S-100 and either tubulin or MAPs was visualized by means of avidin-peroxidase, after SDS-PAGE of the complex mixtures and transfer of the separated proteins onto nitrocellulose. The interaction between S-100 and tubulin was strictly Ca2+ dependent, and resistant to high concentrations of KCl, colchicine, or vinblastine.
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PMID:Molecular interaction of S-100 proteins with microtubule proteins in vitro. 266 85

Mouse antibodies are increasingly used as primary antibodies for immunocytochemistry as more mouse monoclonal antibodies are being produced. The localisation of these antibodies by the PAP technique requires mouse antiperoxidase antibody. A monoclonal antiperoxidase would obviate the limitations of production of a polyclonal mouse antiperoxidase. This paper describes the development of a mouse hybridoma producing such an antibody (MAP A6-2) and the use of this antibody to localise a number of mouse primary antibodies by the PAP technique for both light and electron microscopy. The antibodies localised include monoclonal antienkephalin and antityrosine hydroxylase. MAP A6-2 had a higher affinity in immuno-diffusion experiments and gives slightly better staining with an horse radish peroxidase of a different type from that used for immunisation. Staining was optimum with horse radish peroxidase type X whereas horse radish peroxidase type VI was used for immunisation. Also described is the production of a HAT sensitive variant cell line allowing the possibility of using this hybridoma as a parent cell line for the production of hybrid hybridomas secreting bi-specific antibodies.
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PMID:Development of a mouse antiperoxidase secreting hybridoma for use in the production of a mouse PAP complex for immunocytochemistry and as a parent cell line in the development of hybrid hybridomas. 393 12

Little is known about the microvascular effects of blood replacement solutions. This study was undertaken to develop an animal model suitable for studies of the microcirculatory effects of such solutions and to investigate microvascular responses to isovolemic transfusion with stroma-free hemoglobin (SFH), whole donor blood, or a new potential blood substitute solution containing oxypolyhemoglobin (OPH) as an oxygen carrier. Hamster livers were exposed and the microcirculation studied using intravital epifluorescent video microscopy. 33% blood volume replacement with SFH elevated systemic blood pressure by 25 Torr. Accompanying this increase in pressure was a 36% decrease in sinusoidal blood flow velocity and a 10% decrease in terminal hepatic venular diameters. Terminal portal venular diameters did not change. Decrease in liver sinusoidal perfusion was not due to neutrophil mediated injury, as myeloperoxidase activity in jejunum, liver, kidney, and lung remained unchanged. The reduction in perfusion was likely due to systemic vasoconstriction produced by SFH. In contrast, transfusion with whole blood did not change any of the measured parameters showing the excellent stability of the model. OPH transfused animals exhibited only a small 10 Torr transient increase in MAP 15 min post-transfusion. By 30 min MAP returned to the pre-infusion value. No significant changes were observed in either venular diameters or sinusoidal velocities in this group of animals. These results demonstrate suitability of this model for studies of the microcirculatory and hemodynamic effects of blood replacement solutions. Furthermore, OPH solution produced only minor transient disturbances in microvascular and systemic parameters.
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PMID:Systemic hemodynamic and hepatic microvascular responses to a 33% blood volume exchange with whole blood, stroma-free hemoglobin, and oxypolyhemoglobin solutions. 826 May 79

We investigated the activation of three subfamilies of mitogen-activated protein kinases (MAP kinase), the extracellular regulated kinase (ERK1/2), p38, and c-Jun N-terminal kinase (JNK), by the myeloperoxidase-derived oxidant HOCl, in human umbilical vein endothelial cells (HUVEC) and human skin fibroblasts. Treatment of fibroblasts with 10-30 microM HOCl induced a dose-dependent increase in the tyrosine phosphorylation of several proteins. ERK1/2 was activated by exposure to sublethal concentrations of reagent HOCl or by HOCl generated by myeloperoxidase as shown by immune complex kinase assays. Maximum activation was seen at 20 microM and peak activation occurred within 10 min. Western blot analysis demonstrated activation of p38 with 30 microM HOCl, occurring at 15-30 min. No activation of JNK was detected in the concentration range investigated. These results show that HOCl is able to activate MAP kinases. Effective doses were considerably lower than with H2O2 and the lack of JNK activation contrasts with the activation frequently seen with H2O2. Exposure to HOCl caused a loss of viability in HUVEC that was markedly enhanced when ERK1/2 activation was inhibited by U0126. This suggests that the activation of ERK promotes cell survival in response to the oxidative challenge.
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PMID:Hypochlorous acid stimulation of the mitogen-activated protein kinase pathway enhances cell survival. 1156 22

Hemorrhagic shock (HS) elicits an inflammatory response characterized by increased cytokine production and recruitment of PMN which we previously found to be iNOS dependent. In this study we attempted to remove excess induced-NO by administration of the NO scavenger, NOX, with the goal of suppressing proinflammatory signaling and reducing organ damage. Rats subjected to HS (MAP = 40 mmHg for 100 min) followed by resuscitation and examined 24 h later demonstrated histological signs of lung injury including pulmonary edema as well as an 8.6-fold increase in MPO-positive PMN. These events were accompanied by a 3.9-fold increase in mRNA levels for IL-6, 3.7-fold for ICAM-1, 3.5-fold for IL-1beta, and 7.3-fold for TNFalpha compared to sham animals. Immunostaining of the lungs of shock animals demonstrated IL-6 protein localized to cells lining the luminal sides of bronchiols. These animals also demonstrated a 2-fold and 5.5-fold increase in activation of NF-kappaB and Stat3 (an IL-6 signaling intermediate), respectively. Administration of NOX (30 mg/kg/h beginning at 60 min of shock for total of 4.5 h) resulted in reduced lung injury as measured by a 46% reduction in PMN infiltration, a 20% decrease in wet-to-dry ratio, and improved arterial blood gases. NOX reduced proinflammatory signaling in the lung as demonstrated by a 62% decrease in NF-kappaB binding, 47% reduction in Stat3 binding, a reduction in mRNA expression of 48% for IL-6, 57% for ICAM-1, 67% for IL-1beta, and 64% for TNFalpha, as well as a marked reduction in the intensity of IL-6 protein staining. These data indicate that NOX prevents lung injury in this HS model, possibly through downmodulation of proinflammatory signaling and the shock-induced inflammatory response.
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PMID:A nitric oxide scavenger protects against pulmonary inflammation following hemorrhagic shock. 1183 96

The objective of the present investigation was to compare the antioxidant effect of different forms of Vitamin E (DL-alpha-tocopherol, Mixed Tocopherols, Ronoxan MAP and alpha-tocopherol acetate) and of topical formulations containing these active pharmaceutical ingredients, using chemiluminescence and the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays. Inhibition of the intensity of chemiluminescence, using the H2O2-luminol-horseradish peroxidase (HRP) enzyme system, was measured for 10 min at room temperature in 10 microl samples of each vitamin at different concentrations, and of formulations containing these vitamins. H-donor ability in the DPPH assay, was measured in 10 microl samples at different concentrations of each vitamin, as well as in formulation in ethanol solution; the decrease of absorbency was read at 517 nm. DL-alpha-tocopherol, Mixed Tocopherols and Ronoxan MAP alone or in formulations, markedly inhibited chemiluminescence intensity and decreased absorbency in the DPPH assay in a concentration-dependent manner. Alpha-tocopherol acetate and formulations containing this vitamin did not show antioxidant activity in either assay. Other components of the formulations did not interfere with the measurements, indicating that the methods employed can be used to evaluate antioxidant activity in topical formulations.
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PMID:Comparison of antioxidant activities of tocopherols alone and in pharmaceutical formulations. 1292 91

To assess the potential therapeutic effect of propofol in the treatment of endotoxemia, 76 rats were randomly assigned to 5 groups: control group(A), endotoxemic group(B), pre-treatment group(C), simultaneous treatment group(D) and post-treatment group(E). Five h after endotoxin injection, PO2, pH, MAP, plasma concentrations of Nitrite/nitrate (NO2-/NO3-) and mortality rates were assessed in each group. After the rats were sacrificed, lung tissue was sampled to measure myeloperoxidase (MPO) activity and tumor necrosis factor (TNF)-alpha contents. It was found that endotoxin injection produced progressive hypotension, metabolic acidosis, and a large increase in the plasma NO2-/NO3- concentrations and increased mortality rates in 5 h. Endotoxin injection significantly increased MPO activity and TNF-alpha contents in lung tissue (P < 0.01 or P < 0.05). These changes response to endotoxin were significantly attenuated in the groups B, C and D. But these beneficial effects were blunted in the group E. The results suggest that propofol administration may offer advantages in endotoxemia.
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PMID:Therapeutic effect of propofol in the treatment of endotoxin-induced shock in rats. 1452 47

The study was designed to investigate the effect of progesterone and its gender based variation on myocardial ischemia/reperfusion (I/R) injury in rats. Adult Sprague Dawley rats were divided into vehicle treated reperfusion injury group male (I/R-M), female (I/R-F), ovariectomised (I/R-OVR) and progesterone treatment (I/R-M+PG, I/R-F+PG, I/R-OVR+PG) groups, respectively. I/R injury was produced by occluding the left descending coronary artery (LCA) for 1 h and followed by re-opening for 1 h. Progesterone (2 mg kg(-1) i.p.) was administered 30 min after induction of ischemia. Hemodynamic parameters (+/-dp/dt, MAP), heart rate, ST-segment elevation and occurrence of ventricular tachycardia (VT) were measured during the I/R period. The myocardial infarct area, oxidative stress markers, activities of myeloperoxidase (MPO) and creatine kinase (CK) were determined after the experiment along with the assessment of the effect on apoptotic activity by using DNA fragmentation analysis. Histological observations were carried out on heart tissue. Treatment with progesterone significantly (P<0.05) reduced infarct area, lipid peroxidation (LPO) level and activity of MPO in females (I/R-F+PG) as compared to ischemic females (I/R-F). Progesterone significantly (P<0.001, P<0.05) inhibited serum CK activity and incidences of VT in female rats. Superoxide dismutase (SOD) activity, reduced glutathione (GSH) levels were significantly elevated (P<0.05) in I/R-F+PG group. Internucleosomal DNA fragmentation was less in I/R-F+PG group when compared to I/R-F group. The ischemic male and ovariectomised (I/R-M and I/R-OVR) counterparts did not show any significant change after progesterone treatment. In conclusion, the cardioprotective effect of progesterone on myocardial I/R injury induced damage is based on gender of the animal. The protective effect could be mediated by attenuation of inflammation and its possible interaction with endogenous estrogen.
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PMID:Gender specific effect of progesterone on myocardial ischemia/reperfusion injury in rats. 1758 47


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