Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.2.1.36 (hyaluronidase)
 4,606 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

An enzymic method is described which allows the isolation under comparable conditions of crypt and villus cells from rat jejunum with normal morphologic appearance and high metabolic activity when compared with previous preparations. The method is based on a differential scraping of short lengths of everted small intestine to yield two villus cell fractions and a gut wall residue. The scrapings and the gut tube are incubated for the same length of time in a HEPES-buffered modified Hanks' balanced salt solution containing hyaluronidase, DNase, and soybean trypsin inhibitor. The cells of the crypt region are recovered by a further scraping of the digested gut wall. Cells from all fractions are dispersed by gentle agitation, washed, and harvested by centrifugation. The final crypt and villus cells are 95--99% viable by dye exclusion and exhibit 5--20% cross-contamination on the basis of differential marker enzymes. The isolated crypt and villus cells prepared by the new procedure are suitable for comparative studies of metabolic activity in the absence of chelation-induced structural and metabolic abnormalities.
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PMID:Improved isolation of villus and crypt cells from rat small intestinal mucosa. 49 96

A method is described for the preparation of high yields of viable, dissociated cells from porcine theca interna and corpus luteum and from human and bovine endometrium. The tissues were dissociated by incubation at 37 degrees C in a mixture of 0.5% collagenase, 0.1% hyaluronidase and 0.1% pronase in balanced salt solution containing 1% chicken serum. This procedure consistently provided high yields of structurally and metabolically intact dispersed cells after a digestion period of 60 min. The procedure is superior to methods previously reported in the literature.
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PMID:Enzymatic dissociation of ovarian and uterine tissues. 609 70

A simple method of removing fibroblasts from cultured mammary epithelial cells is described. Primary cultures of both fibroblasts and epithelial cells have been prepared from rat mammary tissue dissociated with collagenase and hyaluronidase. Fibroblasts present as contaminants in the epithelial cell cultures have been selectively removed by incubating cultures at 37 degrees C in Hanks' balanced salt solution that contained antibiotics (100 micrograms/ml) and fungizone (5 micrograms/ml, a treatment which does not appear to decrease cell viability.
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PMID:A simple method for the removal of contaminating fibroblasts from cultures of rat mammary epithelial cells. 664 Jun 76

Instillation of sodium hyaluronate into the anterior chambers of enucleated human eyes caused a 65% decrease in outflow facility (from 0.33 +/- 0.16 microliters/min/mm Hg to 0.08 +/- 0.02 microliters/min/mm Hg). Vigorous anterior chamber irrigation, performed either immediately or three hours after introduction of the sodium hyaluronate, failed to relieve this obstruction. However, irrigation with hyaluronidase restored the facility values to baseline. Tying limbal or corneal 9-0 nylon sutures (for example, in cataract surgery), followed by instillation of sodium hyaluronate into the anterior chamber and subsequent irrigation, produced an overall decrease of 76% in outflow facility (final outflow values were 0.08 +/- 0.03 microliters/min/mm Hg in eyes with corneal wounds and 0.08 +/- 0.04 microliter/min/mm Hg in eyes with limbal wounds). Postoperative intraocular pressure should be monitored closely when sodium hyaluronate is used in cataract surgery. Irrigating the anterior chamber with balanced salt solution after using sodium hyaluronate does not eliminate the possibility of severe postoperative glaucoma.
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PMID:Obstruction of aqueous outflow by sodium hyaluronate in enucleated human eyes. 684 58

Conventional IVF as well as several assisted microfertilization techniques have shown limited success in the horse. After recent positive results achieved with intracytoplasmic injection of a single spermatozoon (ICSI) in human IVF, we chose to try the method in the horse. We compared conventional IVF to ICSI by fertilization rates of oocytes with compact and expanded cumuli and by developmental potential of the resulting embryos. Cumulus-oocyte complexes (COCs) were obtained by aspirating the follicular fluid from the ovaries of slaughtered mares. Complexes showing complete cumulus investment, either compact or expanded, were randomly assigned to IVF or ICSI trials and separately cultured for IVM. Frozen-thawed stallion spermatozoa were prepared for IVF with a swim-up procedure conducted in Talp-Hepes with heparin or for ICSI in Earle's balanced salt solution (EBSS) supplemented with human serum albumin (HSA). Oocytes for IVF were partially decumulated by pipetting, whereas those for ICSI were totally denuded with 80 UI/ml hyaluronidase. Oocytes were fixed, stained and examined for signs of fertilization the day after IVF or ICSI. The percentage of normally fertilized oocytes showing 2 pronuclei or cleavage was significantly higher with ICSI than IVF (29.8%, 17/57 vs 8.7%, 9/103 ; P < 0.01). Significantly higher fertilization rates were observed in oocytes retrieved with an expanded cumulus when submitted to ICSI procedure as compared with IVF (52.2%, 12/23 vs 17.1%, 6 35 ; P < 0.01), whereas in oocytes recovered with a compact cumulus, fertilization rates were low (14.7%, 5/34 with ICSI and 4.4%, 3 68 with IVF; NS). Embryonal development did not occur after culture following IVF, as indicated by absence of cleavage in any of the 93 inseminated oocytes. Following ICSI, 7 of 55 injected oocytes cleaved, 5 of which had shown expanded cumuli; of the 5, 2 were at the 16-cell stage and one each at the 8-, 3- and 2-cell stage, respectively. The other 2 fertilized oocytes, originating from compact cumuli, reached 4- and 8- cell stages, respectively. These results indicate that ICSI can be applied successfully to in-vitro matured equine oocytes to increase the fertilization rates. In addition, it seems that in vitro cytoplasmic maturation of oocytes issuing from a compact cumulus may not be complete enough to lead to a successful fertilization and that ICSI may be a tool to evaluate ooplasmic maturation.
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PMID:Intracytoplasmic sperm injection (ICSI) versus conventional IVF on abattoir-derived and in vitro-matured equine oocytes. 1672 64