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Query: UNIPROT:P30536 (PBS)
 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Rabbit morulae were exposed to a vitrification solution-modified PBS [PB1] medium containing 40% ethylene glycol + 18% Ficoll + 0.3 M sucrose (EFS) for 2, 5, or 10 min at 20 degrees C and were vitrified in liquid nitrogen. When morulae were rapidly warmed, 96% had an intact zona pellucida. When embryos were cultured after removal of the mucin coat, high proportions of them formed blastocoel (79-100%), but the percentage of embryos developed to fully expanded blastocysts decreased with increased exposure time 87%, 40%, and 17%). The survival rate of morulae vitrified after removal of the mucin coat was lower than that of mucin-intact embryos. To assess the development potential in vivo, 131 embryos were vitrified after 2 min of exposure to EFS solution; all the embryos were recovered and 120 were transferred to recipients without removal of the mucin coat, resulting in 78 (65%) full-term fetuses or young. This simple method, which yields high survival both in vitro and in vivo, will be of practical use for vitrifying rabbit embryos.
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PMID:High survival of rabbit morulae after vitrification in an ethylene glycol-based solution by a simple method. 139 2

Mouse morulae were exposed in one step to a vitrification solution (EFS, a modified PBS containing 40% ethylene glycol, 18% Ficoll, and 0.3-M sucrose) at various temperatures, then cooled rapidly in liquid nitrogen, and then warmed rapidly. All of the embryos exposed to the EFS solution for 0.5 min at 25 degrees C before vitrification developed in culture. However, survival rates were lower if the duration of exposure was prolonged to 2, 5, or 10 min. At lower ambient temperatures (20, 10, and 5 degrees C), high survival rates were associated with longer exposure to the EFS solution. The toxicity of the EFS solution was also lower at lower temperatures. The toxic injury of morulae was manifested as decompaction of the blastomeres. Among the three additives in the EFS solution, ethylene glycol, which can cross cell membranes, was responsible for the toxicity. The results show that the optimum time for exposure of the embryos to the EFS solution before rapid cooling varies with the ambient temperature, i.e., 0.5 min at 25 degrees C, 0.5-5 min at 20 degrees C, 2-5 min at 10 degrees C, and 2-10 min at 5 degrees C. If they are exposed for an optimum period, almost all mouse morulae can survive vitrification (94-100%).
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PMID:Survival of mouse morulae vitrified in an ethylene glycol-based solution after exposure to the solution at various temperatures. 149 79

The discovery of glycerol as an effective cryoprotectant for spermatozoa led to research on cryopreservation of embryos. The first successful offspring from frozen-thawed embryos were reported in the mouse and later in other laboratory animals. Subsequently, these techniques were applied to domestic animals. Research in cryopreservation techniques have included studies concerning the type and concentration of cryoprotectant, cooling and freezing rates, seeding and plunging temperatures, thawing temperatures and rates, and methods of cryoprotectant removal. To date, successful results based on pregnancy rates have been obtained with cryopreserved cow, sheep, goat, and horse embryos but no success has been reported in swine. Post-thaw embryo survival has been shown to be dependent on the initial embryo quality, developmental stage, and species. The freezing techniques most frequently used in research and by commercial companies are identified as "equilibrium" cryopreservation. In this technique the embryos are placed in a concentrated glycerol solution (1.4 M in PBS supplemented with BSA) at room temperature and the glycerol is allowed to equilibrate for a 20-min period. During the cooling process the straws are seeded (-4 to -7 degrees C) and cooling is continued at a rate of 0.3 to 0.5 degree C/min to -30 degrees C when bovine embryos may be plunged into LN2. Sheep embryos are successfully frozen with ethylene glycol (1.5 M) or DMSO (1.5 M) rather than with glycerol. Horse embryos have been frozen in 0.5 rather than 0.25 cc straws but with cooling rates and seeding and plunging temperatures similar to those used with bovine embryos. Swine embryos have shown a high sensitivity to temperature and cryoprotectants probably due to their high lipid content and a temperature decrease to 15 or 10 degrees C causes a dramatic increase in the percentage of degenerated embryos. However, a recent study has shown that hatched pig blastocysts survived exposure below 15 degrees C. Recent research has shown that embryos may also be frozen by a "nonequilibrium" method. This rapid freezing by vitrification consists of dehydration of the embryo at room temperature by a very highly concentrated vitrification media (3.5 to 4.0 M) and a very rapid freeze that avoids the formation of ice allowing the solution to change from a liquid to a glassy state. Vitrification solutions consist of combinations of sucrose, glycerol, and propylene glycol. With this technique, 50% pregnancy rates have been reported with the bovine blastocyst.
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PMID:Status of cryopreservation of embryos from domestic animals. 160 26

The purpose of our study was to compare the therapeutic effects of doxorubicin in 3 different formulations: (1) in PBS, (2) in conventional liposomes composed of egg phosphatidylglycerol/egg phosphatidylcholine/cholesterol/dl-alpha tocopherol, and (3) in sterically stabilized, long-circulating "Stealth" liposomes composed of hydrogenated soy phosphatidylcholine/cholesterol/polyethylene glycol-distearoylphosphatidylethanolamine. The doxorubicin formulations were used to treat recently implanted and well-established, growing primary mouse mammary carcinomas, and to inhibit the development of spontaneous metastases from intra-mammary tumor implants. In the treatment of recently implanted primary tumors, the formulations were given in 3 i.v. injections over 15 days, starting 3 or 10 days after tumor implantation. In the treatment of well-established primary tumors, the mice received 4 i.v. injections over 22 days, starting an average 38 days after tumor implantation. In the preventive treatment against metastases, the formulations were given in 4 i.v. injections over 22 days, starting 22 days or 58 days after primary tumor implantation. The Stealth liposome formulation was significantly more effective than the conventional liposome formulation or the free drug in reducing the incidence of metastases from intra-mammary implants of tumor MC19 and tumor MC65, in curing mice with recent implants of tumor MC2A, tumor MC2B, and tumor MC65, and in increasing the 8-week survival of mice with well-established implants of tumor MC2B. It is concluded that the long circulation time of the Stealth liposome doxorubicin formulation accounts for its superior therapeutic effectiveness.
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PMID:Therapy of primary and metastatic mouse mammary carcinomas with doxorubicin encapsulated in long circulating liposomes. 163 42

Polyethylene glycol (PEG) is an excellent embedding medium for immunohistochemical studies. It provides structural preservation superior to frozen sections and increased sensitivity of antigen detection compared with paraffin sections. One limitation of PEG embedment is that PEG sections are difficult to handle and adhere poorly to glass slides. Here we present a simple and effective method for embedding tissues in PEG and transferring the resultant sections onto silanated glass slides. In addition, a method for silver enhanced colloidal gold immunostaining was combined with common dye staining to demonstrate the excellent structure preservation and sensitive antigen detection. Bovine chorionic membrane was fixed with Bouin's fixative, embedded in polyethylene glycol (PEG) 1500, cut into 5-microns sections, flattened over agarose blocks (10 x 10 x 2 mm3), and blotted onto Digene silanated slides. Slides were then washed in PBS, which removed the PEG and agarose blocks. Tissue sections were immunocytochemically stained with dilute antiserum raised in a rabbit against purified bovine placental retinol binding protein (bpRBP). Sections were washed and incubated with 1-nm colloidal gold-labeled goat anti-rabbit IgG. The immunogold particles were enhanced by silver staining (IGSS). Specimens were observed and photographed with an Olympus epipolarization microscope. The new method offered excellent morphological preservation of cell structure and the epipolarization microscopy provided high sensitivity for detection of specific immunogold-silver particles.
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PMID:A new method for transfer of polyethylene glycol-embedded tissue sections to silanated slides for immunocytochemistry. 200 76

A sensitive enzyme-linked immunosorbent assay (ELISA) to measure antibodies against capsular polysaccharide was developed, based on the enhanced binding of polysaccharide to polystyrene microtitration plates. The wells of the microtitration plate were primed with an adipic acid dihydrazide derivative of bovine serum albumin (AH-BSA) (100 micrograms/mL, 0.01 M NaPO4-0.14 M NaCl, pH 7.2 (PBS]. Capsular polysaccharide, the glucuronoxylomannan of Cryptococcus neoformans serotype A, was oxidized with NaIO4 for 5 min; the reaction was then quenched with ethylene glycol. The partially oxidized polysaccharide was dialyzed vs. PBS, and its concentration was adjusted to 50 micrograms/mL with PBS. This solution (100 microL/well) was covalently bound to the AH-BSA primed microtitration plates through formation of a Schiff base between the hydrazide group on the AH-BSA and the aldehyde groups on the polysaccharide. Antimouse IgG-alkaline phosphatase conjugate was used in an indirect ELISA to measure captured murine monoclonal antibodies directed against glucuronoxylomannan. Mean absorbances, after 15 min, were 0.13 in negative control wells, and greater than 0.7 in test wells. No intermediate steps were required to block nonspecific binding of antibody.
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PMID:Enhanced binding of capsular polysaccharides of Cryptococcus neoformans to polystyrene microtitration plates for enzyme-linked immunosorbent assay. 306 47

In 2 radioimmunoassays in use to detect antibodies to dsDNA, the Farr assay and the PEG assay, we observed inhibitory effects of normal human serum (NHS) on the DNA binding by SLE sera. This was found to be due by the fact that, during incubation at 37 degrees C, CO2, introduced in the incubation mixture by the serum, evaporates from the mixture. This results in increase in pH to values well above pH 8.0, which in turn leads to a decreased DNA binding by antibody. When SLE sera are tested at low dilution, this phenomenon may lead to false negative results. Proper pH control, by the use of buffers with a greater buffering capacity than PBS, completely prevented the observed inhibitory effects. However, under these conditions NHS bound significant amounts of DNA in both assays. The non-specific DNA binding by NHS was found to be heat-stable, but could be eliminated either by aerosil treatment of the sera or by addition of dextran sulphate to the incubation mixture. Lipoproteins and, to a lesser extent, the complement component C1q appear responsible for this non-specific binding. To avoid false negative results with SLE sera as well as non-specific binding by NHS, we propose the use of stronger buffers in combination with added dextran sulphate to the incubation mixture in both the Farr assay and the PEG assay.
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PMID:Influence of pH on the detection of low- and high-avidity anti-dsDNA. 618 61

With the immunofluorescence technique (IFT) using Crithidia luciliae as a substrate, 14,417 sera sent to our laboratory for routine anti-dsDNA determination, were screened for the presence of antibodies to dsDNA. The 1,260 sera that were found IFT positive were then assayed with the Farr radioimmunoassay, in which 3H-labelled PM2-DNA is used as antigen. Only 470 sera (37%) were found to be Farr positive. This discrepancy is, at least partially, caused by the fact that the Farr assay does not detect anti-DNA of low avidity, whereas the Crithidia-IFT does. Sixty-eight percent of the IFT-positive/Farr negative sera were found positive with the PEG assay, a radioimmunoassay that also employs double stranded PM2-DNA as antigen, and that also detects anti-dsDNA of low avidity. The IFT performed on IFT positive/Farr negative sera was found to be rather irreproducible. It was shown that this was due to local increases of the salt concentration resulting from the way the assay was performed. The problem could be overcome by careful control of the assay conditions, i.e. never letting Crithidia slides dry up after washing with PBS. In the PEG assay, these sera sometimes showed a DNA binding that decreased with time. It could be shown that this is caused by a parallel increase in pH during the incubation as a result of CO2 evaporation from the serum.
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PMID:Measurement of low avidity anti-dsDNA by the Crithidia luciliae test and the PEG assay. 675 23

Circulating immune complexes (CIC) were first measured in lepromatous patients (LL) by the 125I-C1q binding assay and the polyethylene glycol (PEG) precipitation test. High levels were found by both methods (95 and 90% of positives, respectively). LL-CIC were investigated for the presence of neural antigens. CIC were precipitated in 3.5% PEG, filtered through protein A-Sepharose affinity chromatography, eluted with glycine-HCl, pH 2.8, and washed with PBS; fractions after CIC dissociation were studied by SDS-PAGE and Western blotting. The LL-CIC PEG precipitates and the glycine-HCl eluates were positive in 76 and 71% respectively against anti-myelin basic proteins (MBP) monoclonal antibody, showing a single band at 15-25 kDa similar to the one obtained incubating MBP with anti-MBP. No reaction was detected with CIC-PBS fractions; strips were incubated with other anti-neural antibodies such as anti-glial fibrillary acidic proteins, anti-S-100, and anti-neurofilaments, without any reactivity. Our results demonstrate that LL-CIC contain MBP as an antigen; its significance could be related to the pathogenesis of leprosy since the liberation of MBP after Mycobacterium leprae nerve damage may elicit anti-MBP autoantibodies to myelin breakdown, which reacts with peripheral nerve MBP inducing CIC formation. This mechanism may be important in demyelination and destruction of nerve in leprosy.
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PMID:Identification of myelin basic proteins in circulating immune complexes associated with lepromatous leprosy. 751 Oct 83

A total of 410 goat embryos were divided at random into 9 groups. Cryoprotectants (glycerol, ethylene glycol or dimethylsulfoxide) were added by a 3-step procedure using increasing concentrations of cryoprotectant (0.5 M; 1 M; 1.5 M) in PBS at 10 min intervals. After freezing and thawing, each cryoprotectant was removed by 3 methods: the classic 3-step procedure (cryoprotectant 1 M-10 min; 0.5 M-10 min; PBS alone-10 min); the same procedure, but with sucrose added to the first 2 steps (sucrose 0.25 M and cryoprotectant 1 M-10 min; sucrose 0.25 M and cryoprotectant 0.5 M-10 min; PBS alone-10 min); and a 2-step procedure with sucrose alone (sucrose 0.25 M-10 min; PBS alone-10 min). Each removal protocol was performed for embryos in each cryoprotectant. The viability of the embryo was evaluated by its capacity to subsequently develop during 48 h in vitro culture. For morulae the development rate of the embryos was significantly higher when they were frozen with ethylene glycol than when dimethylsulfoxide or glycerol was used (P < 0.05). For blastocysts the development rate was the same whether they had been frozen with ethylene glycol or dimethylsulfoxide, and was significantly lower when they were frozen with glycerol (P < 0.05). Among the 3 removal procedures tested, the 3-step procedure with sucrose gave the best development rate and differed significantly from the classic 3-step procedure (P < 0.05).
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PMID:Evaluation of cryopreservation techniques for goat embryos. 754 28


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