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 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

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

Pharmacological studies carried out on protoscoleces in vitro to standardize conditions that would permit a preliminary estimate of the efficacy of drugs with potential activity against Echinococcus granulosus are reported. Media such as PBS and Hanks solution, maintenance temperature, different pH values and concentrations of various solvents have been tested to check the effects on protoscolex survival in tubes in vitro. Mebendazole has been used as the pharmacological standard reference. Changes in the viability of protoscoleces have been used to demonstrate pharmacological activity. Best conditions were obtained employing Hanks solution and propylene glycol at low concentrations. Mebendazole was not completely effective at the concentrations achievable in human therapy. Linear, reproducible results demonstrated that Hanks solution provides an ideal medium for pharmacological studies. Among tested solvents, propylene glycol and dimethyl sulphoxide showed no lethal activity at low concentrations. At concentrations similar to those normally obtained in human sera, mebendazole, as in vivo, demonstrated only partial lethality for protoscoleces. The present study represents a new experimental approach to chemotherapy of hydatid disease.
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PMID:An experimental in vitro model for evaluating drugs against protoscoleces of Echinococcus granulosus. 228 76

Four experiments were conducted to determine the composition of a solution suitable for vitrification of immature bovine oocytes. The osmotic and cytotoxic effect of different concentrations (0, 0.25, 0.5 and 1 mol l-1) of nonpermeating solutes (sucrose versus trehalose) were examined. In addition, the effect of permeating cryoprotectants such as glycerol, dimethyl sulfoxide, and propylene glycol (40% w/v) on the viability of oocytes was studied to determine the optimal time of exposure and the most suitable cryoprotectant. Exposure of bovine oocytes to trehalose was less harmful than exposure to sucrose (P < 0.01), and high normospermic fertilization (70%) was achieved after exposure to 0.25 mol trehalose l-1. Propylene glycol was chosen as the cryoprotectant for the vitrification of immature bovine oocytes because of its fast permeating rate and its low cytotoxic effect. The composition of this solution (40% propylene glycol and 0.25 mol trehalose l-1 in PBS containing 4% w/v BSA) appeared to be suitable for vitrification, as the fertilization rate of the vitrified oocytes was 37% (36 of 97).
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PMID:Osmotic and cytotoxic study of vitrification of immature bovine oocytes. 810 16

Seventeen horse embryos recovered on the sixth day after spontaneous ovulation were; 1) washed in PBS (n = 6), 2) treated with 1.5 M 1-2 propanediol (n = 6) or, 3) frozen and thawed using 1.5 M propanediol as the cryoprotectant (n = 5). After treatment, the embryos were incubated for 6 h in medium before they were fixed, serially sectioned and examined microscopically to count the total numbers of interphase, mitotic and pycnotic nuclei. Significant differences were measured only in the mean proportions of pycnotic cells (+/- s.d.), both between the control (9.2 +/- 7.3%) and frozen-thawed embryos (52.8 +/- 37.1%; P<0.05) and between the propanediol-treated (10.8 +/- 4.6%) and the frozen-thawed embryos (P<0.01). Propanediol appears to be minimally toxic to equine embryos but it is a poor cryoprotectant.
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PMID:The effect of propanediol on the morphology of fresh and frozen equine embryos. 959 35

The primary goal of this work was to evaluate the long-term constant zero-order release of progesterone from a waterborne, in situ-gelling, injectable material. The motivation for this is to develop an intrafallopian tube embolization system for contraception. Poly(ethylene glycol) diacrylate (PEGDA, 575 g/mol) or poly(propylene glycol) diacrylate (PPODA, 540 g/mol) as a Michael-type addition acceptor was combined with pentaerythritol-tetrakis (3-mercaptopropionate; a Michael-type addition donor) to create a 75 wt.% emulsion solution in 0.1M PBS (pH 7.4 for PEGDA and pH 12 for PPODA) that gels in minutes by the Michael-type reaction to form a hydrophobic solid. Samples, with approximately 5.5 or 25 wt.% progesterone, were formed in Tygon tubing. Samples (1.6 mm x 1.0 cm cylinders) showed constant, partition-controlled release of progesterone for a prolonged period (time dependent on the mass of progesterone). Cylinders with approximately 25 wt.% load of progesterone exhibited constant release (approximately 40 microg per day) for more than 50 days in both the PEGDA and PPODA systems. This type of release is normally associated with preformed hydrophobic matrix systems. In contrast, these in situ-gelling materials reported here can be used to provide zero-order, partition-controlled release of progesterone and enhance the efficiency of an intrafallopian tube embolization system through progesterone release in an injectable, in situ-forming system.
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PMID:Partition-controlled progesterone release from waterborne, in situ-gelling materials. 1507 95

The survival of ovine embryos (morulae and blastocysts) either frozen by a conventional method or vitrified was investigated in culture. In Experiment I, embryos were vitrified using a solution containing 25% propylene glycol and 25% glycerol. A group of embryos (simulated control) was processed without freezing to evaluate the toxicity of the vitrification solution. In Experiment II, embryos were exposed to a solution of PBS containing 10% glycerol and 0.25 M sucrose placed horizontally in a programmable freezer. Automatic seeding was applied at -7 degrees C in 2 positions on straws and cooled at -0.3 degrees C/min to -25 degrees C and then stored in liquid nitrogen. In vitro development rates of vitrified embryos were 12% (morulae) and 19% (blastocysts). Simulated embryos showed a higher rate of survival than embryos cryopreserved by vitrification (67 and 63%, morulae and blastocysts respectively). In conventional cooling, the blastocysts showed the highest viability percentage (67%) of all the experimental groups but these values decreased significantly in morulae (31%). Differences in temperature between straws placed in distinct positions in the freezing chamber and thermic deviation were observed when automatic seeding was applied. Embryo viability differed from 51 to 75% according the relative position of the embryos within the chamber. Survival was higher when automatic seeding was applied on the meniscus of the embryo column versus the central point of this column (65 vs 21%). The damage of both cryopreservation methods on zona pellucida integrity (27 and 35% in vitrified and conventionally frozen embryos, respectively) had no effect on the in vitro survival.
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PMID:Sheep embryo cryopreservation by vitrification and conventional freezing. 1672 40

In vitro permeation of hinokitiol (HKL) through hairless mouse skin was investigated using a diffusion cell. Either propylene glycol (PG) or ethanol (EtOH) was used as a vehicle for HKL. After applying the HKL solutions of 0.5%. 1%, 2%, and 5% onto the skin, the amount of HKL transferred through the skin into the receptor solution, phosphate-buffered saline (PBS, pH7.4), was determined at a predetermined time intervals for 18 hr using a high performance chromatography (HPLC). EtOH was more effective than PG in terms of in vitro permeation of HKL. This is possibly because EtOH acts as a permeation enhancer. Another reason would be related to the higher thermodynamic activity of HKL in ethanol. To investigate the effect of an enhancer on the in vitro permeation, oleyl alcohol, 1-dodecyl-2-pyrrolidone (DP), and lauric acid were used as enhancers. Each was added to the HKL solution (1%) so that the concentration of the enhancer was 1%. Among the enhancers, DP was the most effective and it enhanced the permeation of HKL approximately 5-10 times.
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PMID:In vitro permeation study of hinokitiol: effects of vehicles and enhancers. 1819 19

We propose that skin electrical current measurements can be used in vitro to effectively rank aqueous solutions containing surfactants and humectants (the enhancer) contacting the skin, relative to a PBS aqueous solution (the control) contacting the skin, based on their ability to perturb the skin aqueous pores. Specifically, we develop an in vitro ranking metric using the increase in the skin electrical current induced by an enhancer relative to the control. Aqueous contacting solutions containing (i) surfactants [SDS (sodium dodecyl sulfate)] and C(12)E(6) [dodecyl hexa (ethylene oxide)], (ii) humectants (glycerol and propylene glycol), and (iii) a control (PBS) were studied. Utilizing the new in vitro ranking metric, these aqueous contacting solutions were ranked as follows (from the mildest to the harshest): glycerol < propylene glycol < PBS < C(12)E(6) < SDS. In order to further develop this ranking methodology, which can potentially lead to the reduction, or elimination, of costly and time-consuming procedures, such as human and animal testing and trial-and-error screening in vivo, it was important to correlate the findings of the in vitro ranking metric with direct in vivo skin barrier measurements. For this purpose, in vivo soap chamber measurements, including transepidermal water loss, visual skin dryness, and chromameter erythema measurements, were carried out on human volunteers using the aqueous surfactant-humectant solutions described above. The results of these in vivo measurements were found to be consistent with the ranking results obtained using the in vitro ranking metric. To further explore the validity of our model and to verify the skin barrier mitigating effect of glycerol, in vivo soap chamber measurements were carried out for aqueous SDS solutions containing 10 wt% added glycerol. These in vivo measurements support our recent in vitro finding that glycerol reduces the average radius and the pore number density of the skin aqueous pores, such that SDS micelles are hindered from penetrating into the skin and inducing skin barrier perturbation.
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PMID:Ranking of aqueous surfactant-humectant systems based on an analysis of in vitro and in vivo skin barrier perturbation measurements. 1830 74

Melatonin is a hormone responsible for the regulation of circadian and seasonal rhythms. This hormone is synthesised in many tissues in the body including the eye, where it regulates important processes. During the recent years, the role of melatonin in the control of IOP has been investigated and it has been demonstrated that melatonin receptors are present and involved in the dynamics of the aqueous humour. 5-Methoxycarbonylamino-N-acetyltryptamine (5-MCA-NAT) is a selective MT3 melatonin receptor agonist. Topical application of this product produces a clear reduction in intraocular pressure (IOP) in New Zealand white rabbits and in glaucomatous monkeys. In this work, the potent ocular hypotensive 5-MCA-NAT has been dissolved in excipients used in currently marketed drug formulations. Until now, this melatonin analogue had been dissolved in either DMSO or ethanol neither of which is suitable for ocular topical application in humans. Solubility assays in the different solvents were performed by the observation of the presence of drug crystals under optical microscopy. 5-MCA-NAT was completely dissolved in propylene glycol (PG) and polyethylene glycol 300 (PEG 300) within 24h. Ophthalmic formulations were prepared from different ratios of PG:PBS and the commercialized Systane product. Quantification of 5-MCA-NAT in the vehicles was assessed by HPLC. In vitro cytotoxicity of the formulations was evaluated by the MTT method and in vivo tolerance of 5-MCA-NAT in the solvents was analyzed by biomicroscopy and specular microscopy. Systane and proportions of PG:PBS up to 10% of PG did not show cytotoxicity in human corneal limbal epithelial cells (HCLE). In vivo experiments showed that the higher the ocular tolerance, the less amount of PG present. The ocular hypotensive effect of 5-MCA-NAT dissolved in the new formulations was checked measuring IOP for 8h after instillation of the substance. The best effect lowering IOP was obtained with 5-MCA-NAT dissolved in PG and diluted with PBS (PG 1.43%) in which 5-MCA-NAT produced a reduction of 28.11+/-2.0% and the effect lasted about 7h. In conclusion, new formulations accepted for ocular topical treatments different from DMSO or ethanol were capable of dissolving the melatonin analogue 5-MCA-NAT, preserving its ocular hypotensive ability. Therefore, the use of 5-MCA-NAT may be possible in the treatment of ocular hypertension and glaucoma.
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PMID:Ophthalmic formulations of the intraocular hypotensive melatonin agent 5-MCA-NAT. 1905 82

In the presence of the nonionic alkyloxyethylene surfactant n-dodecylpentaoxyethylene glycol ether (C12E5), the anionic conjugated polyelectrolyte (CPE) poly{1,4-phenylene-[9,9-bis(4-phenoxy-butylsulfonate)]fluorene-2,7-diyl} (PBS-PFP) dissolves in water, leading to a blue shift in fluorescence and dramatic increases in fluorescence quantum yields above the surfactant critical micelle concentration (cmc). No significant changes were seen with a poly(ethylene oxide) of similar size to the surfactant headgroup, confirming that specific surfactant-polyelectrolyte interactions are important. From UV-visible and fluorescence spectroscopy, dynamic light scattering (DLS), small-angle X-ray scattering (SAXS), cryogenic transmission electron microscopy (cryo-TEM), and electrical conductivity, together with our published NMR and small-angle neutron scattering (SANS) results, we provide a coherent model for this behavior in terms of breakup of PBS-PFP clusters through polymer-surfactant association leading to cylindrical aggregates containing isolated polymer chains. This is supported by molecular dynamics simulations, which indicate stable polymer-surfactant structures and also provide indications of the tendency of C12E5 to break up polymer clusters to form these mixed polymer-surfactant aggregates. Radial electron density profiles of the cylindrical cross section obtained from SAXS results reveal the internal structure of such inhomogeneous species. DLS and cryo-TEM results show that at higher surfactant concentrations the micelles start to grow, possibly partially due to formation of long, threadlike species. Other alkyloxyethylene surfactants, together with poly(propylene glycol) and hydrophobically modified poly(ethylene glycol), also solubilize this polymer in water, and it is suggested that this results from a balance between electrostatic (or ion-dipole), hydrophilic, and hydrophobic interactions. There is a small, but significant, dependence of the emission maximum on the local environment.
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PMID:Solubilization of poly{1,4-phenylene-[9,9-bis(4-phenoxy-butylsulfonate)]fluorene-2,7-diyl} in water by nonionic amphiphiles. 1937 13


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