An in vitro micro-volume procedure for rapid measurement of erythrocytic hexose monophosphate shunt activity

Abstract

Abstract-v-I. A radiometric micro-volume procedure for measurement of erythrocytic hexose monophosphate shunt (HMS) activity in intact cells in vitro is described.2. The procedure is rapid, allowing 200 individual HMS determinations in a single experiment of 5 hr duration.3. The procedure is reproducible, yielding HMS activity means insignificantly different (P > 0.05) between replicate experiments. 4. A profile of sodium nitrite-induced HMS stimulation is reported: HMS was elevated 2-fold (P < 0.001) between zero and 2.5 mM NaN0 2 ; HMS elevation was more distinct (7-fold) between 2.5 and 5.0 mM NaNO z ; maximum activity (22-fold) was observed between 10 and 20 mM NaN0 2 ;

8-Aminoquinoline Therapy for Latent Malaria

“…In that year, the author took a junior technical position (as a graduate student) at ET-WRAIR. There, under the supervision of David Davidson, Jr., and Joan Decker-Jackson, he strived to develop an in vitro means of measuring relative hemolytic toxicity among 8-aminoquinolines (66,67,195,196). The author left that post in 1984 without having achieved that technical objective and with a lasting impression of having been the sole meager resource committed to it before or after his tenure in that era.…”

Methylene blue-mediated hexose monophosphate shunt stimulation in human red blood cells in vitro: Independence from intracellular oxidative injury

“…Physiologically balanced salts solution (PBSS) and phosphate buffered salts solution (PBS) were prepared as described previously (Baird et al, 1984).…”

“…Missouri, U.S.A.) were prepared as previously described (Baird et al, 1984). Reagents for tyrosine assay, l-nitroso-2-naphthol (reagent grade, Aldrich Chemical Co.) and nitric acid (Aldrich Chemical Co.), were also prepared as described elsewhere (Waalkes and Udenfriend, 1957).…”

“…Twenty ml whole blood was drawn by venipuncture from a 24-year-old Caucasian male and immediately washed and albuminized as reported previously (Baird et al, 1984). Cycloheximide (Aldrich Chemical Co.) was added to the red cell suspension (O°C) to 0.5 mM final concentration to prevent incorporation of liberated tyrosine into reticulocyte protein (Goldberg and Boches, 1982).…”

“…HMS activity was measured, with correction for glucose carbon recycling through the shunt, as described previously (Baird et al, 1984). To estimate the possible influence of 0.5 mM cycloheximide upon HMS stimulation induced by Methylene Blue or sodium nitrite, cycloheximide-treated red cells were treated separately with these compounds as described above for measurement of proteolysis and HMS activities.…”