ABSTRACT
Using a monoclonal antibody (LIS-4) to an immunosuppressive factor isolated from boar vesicular gland secretion it was determined that this gland secretes a tissue-specific immunosuppressive molecule that is absorbed onto the acrosome of spermatozoa during ejaculation. Absorption of the immunosuppressive molecule onto murine embryos at the 2-, 4-, 8-cell, morula and blastocyst stages in vitro was evaluated by indirect immunofluorescence. In vivo absorption was detected on the zona pellucida of murine embryos obtained from oviducts injected with the immunosuppressive molecule. Immunofluorescence revealed that the immunosuppressive molecule was not absorbed onto murine embryos after solubilization of the zona pellucida. There was no effect of the antibody to the immunosuppressive molecule on the ability of boar spermatozoa to penetrate the porcine zona pellucida.
Subject(s)
Antibodies, Monoclonal/immunology , Semen/immunology , Suppressor Factors, Immunologic/physiology , Animals , Female , Fluorescent Antibody Technique , Male , Mice , Mice, Inbred BALB C , Sperm-Ovum Interactions , Suppressor Factors, Immunologic/immunology , SwineABSTRACT
The activity of the immunosuppressive fraction and proteinase inhibitor isolated from cow follicular fluid was investigated. The immunosuppressive factor was separated from the accompanying proteinase inhibitor by Sepharose 6B and Sephacryl S-200 column chromatography. In vitro, the fraction significantly reduced mitogen-induced lymphocyte proliferation. In vivo, the fraction inhibited mouse plaque formation. However, the factor had no effect on the development of intact or zona free embryos. The active components have a molecular mass of about 110,000.
Subject(s)
Body Fluids/chemistry , Cattle/immunology , Immune Tolerance , Ovarian Follicle/physiology , Protease Inhibitors/isolation & purification , Animals , Cattle/physiology , Chromatography, Gel , Diestrus , Embryonic and Fetal Development/drug effects , Female , Lymphocyte Activation/drug effects , Lymphocyte Subsets/drug effects , Mice , Mice, Inbred BALB C/embryology , Molecular Weight , Protease Inhibitors/pharmacologyABSTRACT
The effects of an immunosuppressive factor (ISF) isolated from boar seminal vesicle fluid on in vitro and in vivo mouse development were investigated. It was found that the zona pellucida of porcine and mouse oocytes pre-incubated with ISF reacted in indirect immunofluorescence with antibodies against ISF. Further results indicated that the boar ISF had no effect on embryo development.
Subject(s)
Embryonic and Fetal Development/immunology , Immunosuppressive Agents/immunology , Prostatic Secretory Proteins , Proteins/immunology , Semen/immunology , Animals , Embryonic and Fetal Development/drug effects , Female , In Vitro Techniques , Male , Mice , Oocytes/immunology , Pregnancy , Proteins/pharmacology , Seminal Plasma Proteins , Seminal Vesicles/metabolism , Swine , Zona Pellucida/immunologyABSTRACT
The occurrence of microorganisms, including their total counts in boar native ejaculates, was investigated in two stages; the objective of this investigation also was to determine contamination after the sperms were treated with diluents containing the antibiotics ampicillin, gentamycin, apramycin, cefoxitin, or antibiotic combinations penicillin + streptomycin, ampicillin + cefoxitin, gentamycin + cefoxitin and ampicillin + gentamycin. The representation of bacterial species and total counts of microbes in 1 ml diluted sperm stored at a temperature of about 18 degrees C were determined in 24, 48 and 72 h after dilution. The microorganisms were cultivated from all native ejaculates. Proteus sp. (63.3%) and Pseudomonas aeruginosa (51.5% of the total number of examined samples) were the most frequent species. The number of contaminated diluted ejaculates ranged from 12.5 to 95.8% in 24 h after dilution, from 12.5 to 98.5% in 48 h and from 16.8 to 95.8% of the total number of examined ejaculates in 72 h. The occurrence of microorganisms correlated mostly with the efficiency spectrum of the antibiotics or their combinations. The average counts of microorganisms in 1 ml of native ejaculate made 2,363,000 in stage I and 1,472,108 in stage II. The highest average counts in 1 ml of diluted sperm were found in ejaculates containing cefoxitin and apramycin. Gentamycin was the most effective antibiotic used as a sole component (average counts of microorganisms CPM in 1 ml were 416 in 24 h, 955 in 48 h and 2260 in 72 h after dilution); ampicillin and gentamycin were the most efficient combination (14--20--21). This combination exerted very good effects also on Proteus sp. and Pseudomonas aeruginosa.
Subject(s)
Anti-Bacterial Agents/pharmacology , Bacteria/drug effects , Semen/microbiology , Swine/microbiology , AnimalsSubject(s)
Metals/blood , Bismuth/blood , Cadmium/blood , Child , Child, Preschool , Cobalt/blood , Copper/blood , Humans , Iron/blood , Lead/blood , Lithium/blood , Manganese/blood , Methods , Nickel/blood , Pica/blood , Solvents , Spectrophotometry , Strontium/blood , Zinc/bloodABSTRACT
The procedure developed by Browett and Moss (1964) for the semi-automatic determination of the lead content of urine has been adapted for the determination of lead in blood. Determinations are normally carried out in duplicate on 2.0 ml. samples of whole blood and the minimum sample size is 0.5 ml. The organic substances present in blood are destroyed by a manual wet-oxidation procedure and the lead is determined colorimetrically as lead dithizonate using a Technicon AutoAnalyzer. The lower limit of detection, expressed as three times the standard deviation of the blank value, is 5 mug. Pb/100 ml. blood. The standard deviation of the method in the upper range of normal blood lead level of 30 mug. Pb/100 ml. blood (Moncrieff, Koumides, Clayton, Patrick, Renwick, and Roberts, 1964), is +/- 3 mug. Pb/100 ml. blood. Ten samples per hour may be estimated in duplicate.
