Animals ; Enzyme Inhibitors ; pharmacology ; Hepatitis ; immunology ; Immune Sera ; pharmacology ; Mice ; Sodium-Potassium-Exchanging ATPase ; antagonists & inhibitors ; metabolism

This study evaluates the pathogenesis of rheumatoid arthritis by producing immune complex induced arthritis with an intra-articular injection of BSA in immunized rabbits, and the effect of systemic administration of cyclophosphamide and local administration of anti-macrophage serum. The reduction of inflammatory reaction by cyclophosphamide administration appears to be caused mainly by selective depletion of the neutrophils, and partly by immune suppression. It appears that the rabbit abdominal macrophage has the common morphologic, functional and antigenic patterns with the M-type synovial lining cells. There is another possibility that the cross-reacting antigens between macrophage and the M-type cell of the synovial lining may exist. It is concluded that in this experimental immune complex arthritis, the site of localization of immune complexes seems to be the synovial, M-type cell, and the tissue injury of synovium is largely mediated not only by neutrophils and complement, but also by macrophages.
Animal ; Antigen-Antibody Complex* ; Arthritis, Rheumatoid/etiology* ; Cyclophosphamide/pharmacology ; Immune Complex Diseases/etiology* ; Immune Sera/pharmacology ; Macrophages ; Rabbits ; Serum Albumin, Bovine/administration & dosage ; Synovial Membrane/pathology

The effects of ethyl alcohol and pig serum administration on the development of preneoplastic hepatic enzyme-altered foci were examined in an in vivo mid-term assay system. Rats were initially given a single dose (200 mg/Kg) intraperitoneal injection of diethylnitrosamine (DEN). Two weeks later, treatment was started with 10% ethanol + 10% sucrose solution, 10% sucrose solution, or tap water as drinking water for 6 weeks with or without intraperitoneal injection of porcine serum twice a week. All rats were subjected to a two-thirds partial hepatectomy at week 3. The modification potentials were evaluated by comparing the number and area per cm2 of glutathione S-transferase placental form-positive (GST-P+) foci in the liver of each group. As a result, ethanol significantly enhanced the development of GST-P+ foci. Unfortunately, the porcine serum injection produced no hepatic fibrosis and no significant alteration in GST-P+ foci.
Animals ; Diethylnitrosamine/*toxicity ; Ethanol/*pharmacology ; Glutathione Transferase/*metabolism ; Immune Sera/pharmacology ; Liver Cirrhosis, Alcoholic/enzymology ; Male ; Placenta/drug effects/*enzymology ; Precancerous Conditions/*chemically induced/enzymology ; Rats ; Rats, Inbred F344 ; Survival Rate ; Swine

AIMTo study the effect of antidigoxin antiserum on oxygen stress induced by myocardial ischemia/reperfusion (MI/R) injury in rats.

METHODSSprauge Dawley rats were submitted to ligate left anterior descending coronary artery 30 min followed by 45 min reperfusion. Experiment animals were randomly divided into seven groups including sham group, MI/R group, normal salina group, verapamil group and three antidigoxin antiserum groups from low to high dose. The left ventricular myocardial tissue sample of ischemia were processed and measured the level of endoxin and malondialdehyde (MDA), the activities of Na+, K(+) -ATPase and superoxin dismutase (SOD). The myocardia morphology was observed.

RESULTSThe levels of endoxin and MDA increased and the activities of Na+, K(+) -ATPase and MDA were inhibited significantly in MI/R and saline groups. Including verapamil group in comparison to MI/R and saline groups, MDA level decreased and SOD activities partly reserved, meanwhile, only in three antidigoxin antiserum groups, the myocardial endoxin level was remarkably decreased, Na+, K(+) -ATPase activities were drastically increased. The myocardial histological morphology was significantly improved.

CONCLUSIONAntidigoxin antiserum, an endoxin mutual clone antibody, had the effect of attenuating the damage of oxygen free radicals induced by MI/R via to antagonizing the inhibition effect of endoxin on myocardial membrane Na+, K(+) -ATPase activities.

Animals ; Cardenolides ; antagonists & inhibitors ; Digoxin ; pharmacology ; Immune Sera ; pharmacology ; Malondialdehyde ; analysis ; Myocardial Reperfusion ; Myocardial Reperfusion Injury ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Saponins ; antagonists & inhibitors ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Superoxide Dismutase ; metabolism

Animals ; Collagen Type IV ; metabolism ; Fibronectins ; metabolism ; Glomerulonephritis, Membranous ; metabolism ; pathology ; Immune Sera ; immunology ; Kidney Glomerulus ; pathology ; ultrastructure ; Male ; Malondialdehyde ; blood ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; pharmacology ; Superoxide Dismutase ; blood ; Taurine ; pharmacology ; Transforming Growth Factor beta1 ; metabolism

This study was aimed to prepare the polyclonal antibody against the soluble proliferation-inducing ligand (sAPRIL) antigen and to investigate its effects in suppressing sAPRIL mediated lymphocyte proliferation. Mutated recombinant sAPRIL protein, which lacks biological activity but maintains immunogenicity, was used as antigen to immunize humanized SCID mice. Sera were obtained at 6 weeks after immunization. Indirect ELISA and Western blot were used to detect the antibody titer and specificity. The inhibition of polyclonal antibodies on Raji and Jurkat cell proliferation stimulated by sAPRIL was assessed by the MTT assay. The results showed that the mutant of sAPRIL could induce the production of polyclonal antibodies against human sAPRIL. Western blot and indirect ELISA analyses indicated that the anti-serum had higher specificity with a titer of 1:640. Functional analysis revealed that these polyclonal antibodies significantly inhibited the proliferation of Raji and Jurkat cell stimulated by sAPRIL (p < 0.05). It is concluded the polyclonal antibody against human sAPRIL is successfully prepared, which can inhibit the proliferation of Raji and Jurkat cells stimulated by sAPRIL in vitro.
Animals ; Antibodies ; genetics ; immunology ; pharmacology ; Antibody Specificity ; immunology ; Cell Proliferation ; drug effects ; Cloning, Molecular ; Humans ; Immune Sera ; analysis ; immunology ; Jurkat Cells ; Mice ; Tumor Necrosis Factor Ligand Superfamily Member 13 ; genetics ; immunology

Prior to fertilization sperm has to undergo an activation process known as capaciation, leading to the acrosome reaction. Till now, little is known about the mechanism for preventing premature capacitation in sperm although decapacitation factors from various sources have been thought to be involved. In this study, we report that NYD-SP27, an isoform of phospholipase C Zeta 1 (PLCZ1), is localized to the sperm acrosome in mouse and human spermatozoa by immunofluorescence using a specific antibody. Western blot and double staining analyses show NYD-SP27 becomes detached from sperm, as they undergo capacitation and acrosome reaction. The absence of HCO3-, a key factor in activating capacitation, from the capacitation-inducing medium prevents the loss of NYD-SP27 from sperm. The anti-NYD-SP27 antibody also prevents the loss of NYD-SP27 from sperm, reduced the number of capacitated sperm, inhibited the acrosome reaction induced by ATP and progesterone, and inhibited agonist-induced PLC-coupled Ca2+ mobilization in sperm, which can be mimicked by the PLC inhibitor, U73122. These data strongly suggest that NYD-SP27 is a physiological inhibitor of PLC that acts as an intrinsic decapacitation factor in sperm to prevent premature capacitation and acrosome reaction.
Acrosome ; drug effects ; metabolism ; Acrosome Reaction ; physiology ; Adult ; Animals ; Calcium ; metabolism ; Fluorescent Antibody Technique, Indirect ; Humans ; Immune Sera ; pharmacology ; Male ; Mice ; Middle Aged ; Phosphoinositide Phospholipase C ; immunology ; metabolism ; Sperm Capacitation ; drug effects ; physiology ; Spermatozoa ; drug effects ; metabolism

OBJECTIVE: To study cellular mechanism of cardiomyocytes injury in the early stage of crush injury by observing some effects of crush injury rat sera on cultured neonatal rat cardiomyocytes. METHODS: One to three days old neonatal rat cardiomyocytes were cultured in vitro and some effects of crush injury rat sera on beating rate, cell surface area, total protein content, 3H-Leu incorporation, intracellular calcium concentration ([Ca2+]i) and Fos protein expression were observed in cultured rat cardiomyocytes. RESULTS: Compared with normal rat serum group, crush injury rat sera decreased beating rate(beats/min) of cardiomyocytes from 88.3 to 26.4, cell surface area, total protein content, 3H-Leu incorporation, [Ca2+]i (nmol/L) and PI of Fos protein expression were increased. CONCLUSION Crush injury rat sera suppress cell beating, increase intracellular calcium, induce Fos protein synthesis and cause cell hypertrophy, which may cause cardiac injury in the early stage of rush injury.
Animals ; Calcium/metabolism* ; Cell Size/drug effects* ; Cells, Cultured ; Disease Models, Animal ; Extremities/injuries* ; Heart Injuries/pathology* ; Heart Rate/drug effects* ; Immune Sera/pharmacology* ; Myocytes, Cardiac/pathology* ; Proto-Oncogene Proteins c-fos/metabolism* ; Rats ; Rats, Sprague-Dawley

An improved and practical synthesis of racemic 11-demethylcalanolide A [(+/-)-1] was developed. This improved process involved Pechmann reaction on phloroglucinol with ethyl butyrylacetate to give 5,7,-dihydroxy4-n-propylcoumarin (3). Poly phosphoric acid (PPA) catalyzed acylation of compound (3) with crotonic acid, then intramolecular cyclization was achieved simultaneously in one step to afford the key intermediate chromanone (4). A microwave assisted synthetic method preparing chromene (6) using chromenynation of chromanone (4) with 1, 1-diethoxy-methyl-2-butene was conducted. Luche reduction of chromene (6) using NaBH4 with CeCl3 x 7H2O preferably gave (+/-)-1. The overall yield of this four step synthesis of (+/-)-1 was around 32% increasing one fold more than that of the previous method. An in vitro investigation showed that (+/-)-1 exhibited inhibitory activities against both wild-type and drug-resistant HIV-1 in HIV-1 RT and cell culture assay, and significant synergistic effects in combination with AZT, T-20, and indinavir. Its LD50 of acute toxicity in mice by intragastric administration and by intraperitoneal injection were 735.65 mg kg(-1) and 525.10 mg x kg(-1), respectively. The Cmax and AUC(0-infinity) were 0.54 microg x mL(-1) and 1.08 (microg x mL(-1) x h, respectively. The dynamics study of the inhibition of mice sera on HIV-1 RT showed that mice treated with 100 mg x kg(-1 (+/-)-1 once intraperitoneally were similar to that of 5 mg x kg(-1) of known clinical effective anti-HIV-1 drug neverapine. The results suggested that further investigation of the anti-HIV candidate (+/-)-1 was warranted.
Animals ; Anti-HIV Agents ; chemical synthesis ; immunology ; pharmacology ; toxicity ; Drug Synergism ; HIV Reverse Transcriptase ; metabolism ; HIV-1 ; drug effects ; enzymology ; Humans ; Immune Sera ; pharmacology ; Indinavir ; pharmacology ; Lethal Dose 50 ; Male ; Mice ; Pyranocoumarins ; chemical synthesis ; immunology ; pharmacology ; toxicity ; Reverse Transcriptase Inhibitors ; chemical synthesis ; immunology ; pharmacology ; toxicity ; Zidovudine ; pharmacology

To study whether the antibody against the testis form of the nuclear autoantigenic sperm protein (tNASP) could result in reproductive failure, we successfully cloned and expressed a 339-bp cDNA fragment of mouse tNASP (mtNASP). Using mouse as a model, recombinant mtNASP (rmtNASP) and a synthetic peptide, human tNASP(393-408) (htNASP(393-408)), were investigated for their antifertility effect. Active immunization with rmtNASP or the synthesized peptide raised high antibody titers in the immunized mice. Sperm-egg binding and fusion assay were carried out in 8-10-week-old BALB/c mice. Sperm-egg binding and in vitro fertilization of mouse oocytes were inhibited by co-incubation of zona-free mouse oocytes with capacitated mouse spermatozoa in the presence of varying concentrations of the antisera against rmtNASP. There was a significant antifertility effect in animals immunized with rmtNASP or the synthesized peptide. The effect on fertility in the mice immunized with the synthesized peptide was reversible. Our data indicate that active immunization with rmtNASP antigen may induce a strong antibody response that causes an inhibition of fertility.
Adult ; Animals ; Autoantibodies ; administration & dosage ; immunology ; Autoantigens ; chemistry ; immunology ; pharmacology ; Contraception, Immunologic ; Female ; Fertility ; drug effects ; immunology ; Humans ; Immune Sera ; immunology ; pharmacology ; Male ; Mice ; Nuclear Proteins ; chemistry ; immunology ; pharmacology ; Rabbits ; Recombinant Proteins ; immunology ; Sequence Analysis, Protein ; Sperm Motility ; drug effects ; immunology ; Sperm-Ovum Interactions ; immunology ; Spermatozoa ; drug effects ; immunology ; Vaccines, Contraceptive ; immunology ; pharmacology