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Briefly, two liters of 24 h culture of S. boulardii were centrifuged at 3.000×g for 10 min. The supernatant was decanted and extracted with one-fifth volume of ethyl acetate with changes of ethyl acetate every half an hour of extraction.
After 1 hour of extraction (on a magnetic stirrer), the solution was centrifuged at 4,000 rpm for 10 minutes.
The aqueous phase, after 1 hour of extraction, containing 12.5 mg L-1 of ribitol as an internal standard, was evaporated over-night in a vacuum concentrator and the tube was then returned to the -80°C freezer until use.
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Samples should be measured within a few hours of extraction.
After 24 hours of extraction the suspension was centrifuged at 2000 g for 30 minutes.
Plasma was separated by centrifugation at 3,000 r.p.m. for 10 minutes within eight hours of extraction and then stored at −20°C.
Diagnostic paracentesis was performed on the bed side by sterile method with a 21 G needle attached to 20 cc syringe, and then collected into Ethylene Diamine Tetra Acetate (EDTA) tube and analyzed within 3 hours of extraction.
Nuclei were transported to our laboratories in vials with gauze moistened with balanced salt solution at room temperature and were placed in primary fixative within 4 hours of extraction.
Mechanical testing was done within 48 hours of tissue extraction.
Smoking within the first 24-48 (This number varies up to 7 days or even 3 weeks, it depends on the individual but at least 24 hours) hours of an extraction can also lead to increased risk of a dry socket.
Day 0 and 8 explants were lyophilized and extracted for 48 hours at 4°C in 10 volumes of extraction buffer (4 M guanidine hydrochloride, 10 mM 2- N-morpholino)ethanesulfonic acid, 50 mM sodium acetate, 5 mM ethylenediaminetetraacetic acid, 5 mM iodoacetic acid at pH 6.5) with protease inhibitors.
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