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Acta Pharmacologica Sinica Apr 2000To evaluate the protective effect of anti-digoxin antiserum on hypoxia-reoxygenation induced injured myocardium and its mechanism.
AIM
To evaluate the protective effect of anti-digoxin antiserum on hypoxia-reoxygenation induced injured myocardium and its mechanism.
METHODS
Anti-digoxin antiserum of different concentrations was used, its effect on endoxin and ATPase activity in cell membrane in hypoxia-reoxygenation myocardium model was observed.
RESULTS
The level of endoxin was remarkably higher, ATPase activities in cell membrane were remarkably lower in hypoxic group and hypoxia-reoxygenation injury group than those of normal group; anti-digoxin antiserum could resume ATPase activity in a concentration-dependent manner.
CONCLUSION
Rise of endoxin was the molecular biological basis of myocardial damage during myocardial hypoxia-reoxygenation. Anti-digoxin antiserum had lessened myocardial injury and had a protective effect on hypoxia-reoxygenation myocardium by antagonizing effect of endoxin.
Topics: Animals; Calcium-Transporting ATPases; Cardenolides; Cell Hypoxia; Cell Membrane; Digoxin; Enzyme Inhibitors; Female; Immune Sera; Male; Myocardial Reperfusion Injury; Myocardium; Rabbits; Saponins
PubMed: 11324464
DOI: No ID Found -
Cancer Biology & Therapy Apr 2013Among the different types of tests used for cancer diagnosis, molecular tests have been increrasingly incorporated because of their ability to detect either expression...
Among the different types of tests used for cancer diagnosis, molecular tests have been increrasingly incorporated because of their ability to detect either expression or functional changes in the molecules associated with the disease. Mammaglobin is a protein found in mammary tissue and can be detected in serum. This protein has been proposed as a biomarker to diagnose breast cancer, given that patients exhibit an increased amount of the protein in serum and tumor tissue, in comparison to healthy individuals. The ELISA test was used in the present study to detect mammaglobin in blood samples from 51 breast cancer patients and 51 control individuals. Antibodies against mamaglobin were generated in rabbits by using the following synthetic peptides: A (amino acids 13 to 21), B (amino acids 31 to 39), C (amino acids 56 to 64) and a D peptide, corresponding to the protein isoform without three amino acids (59, 60 and 61 amino acids) from peptide C. All peptides were immunogenic and allowed generation of antibodies that were able to discriminate patients from controls. The best results were obtained for antiserum B, achieving the best sensitivity (86.3%) and specificity (96%).
Topics: Adult; Aged; Aged, 80 and over; Amino Acid Sequence; Animals; Biomarkers, Tumor; Breast Neoplasms; Case-Control Studies; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immune Sera; Mammaglobin A; Middle Aged; Peptide Fragments; Rabbits
PubMed: 23358476
DOI: 10.4161/cbt.23614 -
The Journal of Neuroscience : the... Dec 1984Nerve growth factor (NGF) is known to be essential for survival and maintenance of sympathetic ganglia and of embryonic sensory ganglia of neural crest origin. The...
Nerve growth factor (NGF) is known to be essential for survival and maintenance of sympathetic ganglia and of embryonic sensory ganglia of neural crest origin. The present study examined the physiological and pharmacological roles of NGF in the postnatal development of sensory neurons in the dorsal root ganglion (DRG). In contrast to what is generally stated in the literature, administration of NGF antiserum to newborn rats for a period of 7 days resulted in a significant (approximately 20%) reduction of neuronal number in the lumbar DRG. Size spectrum analysis of surviving neurons revealed a shift toward larger sizes, presumably due to a preferential loss of small cells. The number of neurons in the L5 DRG was studied at various times after unilateral sciatic nerve crush in 1-day-old rats. Axotomy resulted in a substantial loss (40 to 50%) of neurons in the immature DRG. Administration of NGF antiserum to animals with axotomized DRG did not increase cell death when compared with the axotomized controls. However, the number of neurons in the antiserum-treated ganglia decreased by the same percentage (20%) when compared with the control serum-treated ganglia before and after axotomy. Treatment with NGF initially prevented the loss of neurons in the axotomized DRG. However, some neurons died during the first week despite continued NGF administration; and, subsequent to NGF withdrawal, neuronal number decreased to the same level as in control animals. Thus, removal of exogenous NGF resulted in the death of the sensory neurons which had been maintained.(ABSTRACT TRUNCATED AT 250 WORDS)
Topics: Animals; Axons; Ganglia, Spinal; Growth; Immune Sera; Nerve Growth Factors; Neurons, Afferent; Rats; Rats, Inbred Strains; Spinal Cord Injuries
PubMed: 6502217
DOI: 10.1523/JNEUROSCI.04-12-02986.1984 -
Biological & Pharmaceutical Bulletin Feb 2003We previously showed that enzyme immunoassay (EIA) of beta-methyldigoxin (MDx3) using anti-MDx3 3'-hemisuccinate-bovine serum albumin antiserum (Antiserum-I) was... (Comparative Study)
Comparative Study
We previously showed that enzyme immunoassay (EIA) of beta-methyldigoxin (MDx3) using anti-MDx3 3'-hemisuccinate-bovine serum albumin antiserum (Antiserum-I) was superior to that using commercial anti-digoxin antiserum (Antiserum-II) in terms of specificity and that pretreatment of human serum with phenyl boric acid (PBA) column was effective. In the present study, we examined the precision of EIA using Antiserum-I and the recovery of MDx3 after PBA column treatment in rat serum, and also investigated pharmacokinetic changes of MDx3 in rats. The intra- and inter-assay variations and recovery tests using Antiserum-I were good. The PBA column was effective in selectively separating MDx3 from rat serum containing MDx3 and its metabolites. The recovery tests using Antiserum-I with PBA column showed about 110% and the interference of metabolites of MDx3 was negligible. Serum concentration-time courses of MDx3 by EIA using Antiserum-I with PBA column and Antiserum-I were lower than that using Antiserum-II. The distribution volume at steady state and total body clearance values of MDx3 in these conditions were significantly higher than those using Antiserum-II. The usefulness of PBA column was ascertained, while effects of PBA column on these parameters were not significant. In addition, rapid absorption of MDx3 was observed by EIA using Antiserum-I with PBA column. These results suggest that EIA using Antiserum-I with PBA column for the pretreatment of serum samples should be a more useful and valuable system in therapeutic drug monitoring and pharmacokinetic studies of the unchanged type of MDx3 than Antiserum-II.
Topics: Animals; Immune Sera; Immunoenzyme Techniques; Male; Medigoxin; Rats; Rats, Wistar
PubMed: 12576688
DOI: 10.1248/bpb.26.247 -
British Journal of Cancer Jun 1983CEA was extracted by the perchloric acid method from primary adenocarcinomas of the colon and the ovary, from ascitic and pleural fluids from patients with pancreatic,... (Comparative Study)
Comparative Study
CEA was extracted by the perchloric acid method from primary adenocarcinomas of the colon and the ovary, from ascitic and pleural fluids from patients with pancreatic, lung and breast cancer, and from the cyst fluid of a benign ovarian cystadenoma. Further purification included gel filtration and affinity chromatography. Antisera against CEA from colon, breast, ovary, lung and pancreatic cancer were produced in rabbits. In double diffusion experiments, all these CEA samples showed a reaction of complete immunological identity with all the anti-CEA sera, whatever their origin. CEA from colon, breast, pancreas and ovary were labelled with 125I and used in radioimmunological experiments. In a radioimmunological system where the tracer and the antiserum were constant, all the CEA used as standards gave parallel inhibition curves, having nearly identical slopes. This was another criterion of immunological identity. Sera of numerous cancer patients were assayed in several RIA systems, one of them being the classical system with colonic CEA as tracer and anti-colonic CEA as antiserum, the others being "organ specific" systems. The values obtained in these assays were found to be highly correlated: the rank coefficient of correlation between colonic and breast cancer RIA systems was rs = 0.96, that between colonic and ovarian RIA systems, 0.92, that between colonic and pancreatic RIA systems, 0.97 and that between colonic and lung RIA systems 0.96. It is thus concluded that by use of different organ-derived CEA preparations and their corresponding polyclonal antisera, no significative differences in serum CEA levels may be expected. No evidence of organ specificity of serum CEA was found.
Topics: Carcinoembryonic Antigen; Female; Humans; Immune Sera; Immunodiffusion; Neoplasms; Organ Specificity; Radioimmunoassay
PubMed: 6407507
DOI: 10.1038/bjc.1983.137 -
Infection and Immunity Jan 1981The present study examines a mouse model of infection due to group B Streptococcus serotype III (GBS-III) as to the route and timing of antiserum administration for...
The present study examines a mouse model of infection due to group B Streptococcus serotype III (GBS-III) as to the route and timing of antiserum administration for protection and quantitation of bacteremia with and without antiserum. Data for these parameters are contrasted with those after challenge with serotype Ia of group B Streptococcus (GBS-Ia). An intraperitoneal injection of GBS organisms and protective antiserum from a single syringe can be used to create an animal model of disease. Intraperitoneal injection of GBS-III resulted in bacteremia at 0.5 h both in animals who did not receive antiserum (17.4 X 10(2) +/- 7.6 X 10(2) colony-forming units per ml of blood samples) and in animals who received antiserum (19.3 X 10(1) +/- 6.8 X 10(1) colony-forming units per ml). Although intraperitoneal injection of GBS-Ia also resulted in bacteremia evident by 0.5 h in unprotected animals (30.1 X 10(2) +/- 3.8 X 10(2) colony-forming units per ml), no bacteremia occurred in protected recipients of this organism. Bacteremia due to GBS-Ia and GBS-III logarithmically increased until at least 7 h. Bacteremia due to GBS-III in protected animals was cleared by 24 h. Protection against GBS disease did not require simultaneous or proximate administration of the organism and the antiserum. Mice could be protected from death after intraperitoneal challenge with GBS-III or GBS-Ia by antiserum administered intravenously or intraperitoneally from 6 h before to 2.5 h after challenge.
Topics: Animals; Antibodies, Bacterial; Immune Sera; Injections, Intraperitoneal; Injections, Intravenous; Male; Mice; Sepsis; Streptococcal Infections; Streptococcus agalactiae; Time Factors
PubMed: 7011999
DOI: 10.1128/iai.31.1.391-395.1981 -
Developmental Dynamics : An Official... May 1993The complex topological association of Sertoli cells and spermatogenic cells in the testis suggests the existence of cell surface adhesion molecules that regulate...
The complex topological association of Sertoli cells and spermatogenic cells in the testis suggests the existence of cell surface adhesion molecules that regulate cellular interactions within the seminiferous epithelium. The recent report of N-cadherin mRNA expression in the mouse testis implies the involvement of this known adhesion molecule in testicular cell binding. Accordingly, here we report that (1) N-cadherin is found on the surface membranes of rat spermatogenic cells and on Sertoli cells, and (2) that N-cadherin is a partial mediator of Sertoli cell-germ cell adhesion as tested in an vitro cell-cell binding assay. Antiserum directed against the N-cadherin cell adhesion recognition sequence was used for Western blot anlaysis of purified plasma membranes from Sertoli cells and from spermatogenic cells. Both membrane preparations exhibited reactivity at an appropriate M(r) of about 130 kDa. In addition, immunofluorescence assays demonstrated that both germ cells and Sertoli cells were labeled by anti-N-cadherin. Finally, the antiserum was included in a cytometer-assisted cell-cell binding test to determine its inhibitory ability. The antiserum consistently reduced specific testicular cell-cell adhesion by 30%-50%. This is the first demonstration that antibodies directed against the cadherin cell adhesion recognition sequence are capable of inhibiting cell-cell interactions. Pre-incubation of either rat Sertoli cells or spermatogenic cells alone was sufficient to achieve statistically significant inhibition of intercellular adhesion. We conclude, therefore, that N-cadherin is expressed by both Sertoli cells and spermatogenic cells and that N-cadherin is one of a number of regulatory molecules mediating local cellular associations in the mammalian seminiferous tubule.
Topics: Animals; Blotting, Western; Cadherins; Cell Adhesion; Cell Membrane; Fluorescent Antibody Technique; Germ Cells; Immune Sera; Male; Sertoli Cells; Spermatogenesis
PubMed: 8400407
DOI: 10.1002/aja.1001970102 -
Biological & Pharmaceutical Bulletin May 2004To establish a method for quantitative analysis of HM-1 killer toxin (HM-1), two purified mouse monoclonal antibodies, 1F1 and 4A2, and rabbit polyclonal antiserum... (Comparative Study)
Comparative Study
To establish a method for quantitative analysis of HM-1 killer toxin (HM-1), two purified mouse monoclonal antibodies, 1F1 and 4A2, and rabbit polyclonal antiserum against HM-1 were prepared. Both monoclonal antibodies were classified as IgG1(kappa) subtype, and did not neutralize the killing activity of HM-1. By SPOTs analysis, the epitope of 1F1 was found in the sequence of CDPNTG with a corresponding sequence of 11-16 from N-terminal amino acid residues of HM-1, but the epitope of 4A2 was not determined. Using 4A2 and polyclonal antiserum, the sandwich enzyme-linked immunosorbent assay (ELISA) was applied to establish the quantitative determination of HM-1. The concentration of HM-1 was determined successfully at the range of 2.5-100 ng/ml. But in the case of 1F1, the method was not established. Genes were constructed to apply the system to the measurement of the secreted concentrations of mutant HM-1, and it was evident that the production of mutant toxins varied among HM-1 mutant genes. The findings of this study are unique in determinimg the epitope of monoclonal antibody against HM-1, and in quantifying the HM-1 using the spot analysis and sandwich ELISA methods.
Topics: Animals; Antibodies, Monoclonal; Enzyme-Linked Immunosorbent Assay; Immune Sera; Killer Factors, Yeast; Mice; Mycotoxins; Pichia
PubMed: 15133246
DOI: 10.1248/bpb.27.691 -
Journal of Bacteriology Oct 1963Reiter, Harvard (U.S. Biological Laboratories, Fort Detrick, Frederick, Md.). Effects of antiserum on adsorbed phage. J. Bacteriol. 86:637-641. 1963.-The penetration of...
Reiter, Harvard (U.S. Biological Laboratories, Fort Detrick, Frederick, Md.). Effects of antiserum on adsorbed phage. J. Bacteriol. 86:637-641. 1963.-The penetration of T(3) bacteriophages attached to the host cells could be delayed or entirely prevented by antiserum against T(3) phage. The antiserum-sensitive period of the attached phages increased at lower temperatures. The degree of inhibition increased with antiserum concentration and with early exposure to antiserum. Bacteriophages T(1) and T(4) were not affected by antiserum in this manner; T(7) reacted similarly to T(3).
Topics: Bacteriophages; Coliphages; Immune Sera; Research
PubMed: 14066455
DOI: 10.1128/jb.86.4.637-641.1963 -
Journal of Visualized Experiments : JoVE Feb 2014There are over 90 different capsular serotypes of Streptococcus pneumoniae (the pneumococcus). As well as being a tool for understanding pneumococcal epidemiology,...
There are over 90 different capsular serotypes of Streptococcus pneumoniae (the pneumococcus). As well as being a tool for understanding pneumococcal epidemiology, capsular serotyping can provide useful information for vaccine efficacy and impact studies. The Quellung reaction is the gold standard method for pneumococcal capsular serotyping. The method involves testing a pneumococcal cell suspension with pooled and specific antisera directed against the capsular polysaccharide. The antigen-antibody reactions are observed microscopically. The protocol has three main steps: 1) preparation of a bacterial cell suspension, 2) mixing of cells and antisera on a glass slide, and 3) reading the Quellung reaction using a microscope. The Quellung reaction is reasonably simple to perform and can be applied wherever a suitable microscope and antisera are available.
Topics: Animals; Antibodies, Bacterial; Antibody Specificity; Bacterial Capsules; Horses; Immune Sera; Serotyping; Streptococcus pneumoniae
PubMed: 24637727
DOI: 10.3791/51208