A 30-mL supernatant sample collected from the desynchronized P. falciparum culture and a sample collected from the uninfected RBC culture were concentrated 2-fold by freeze-drying. The resulting suspension was acidified with 200 mL of concentrated hydrochloric acid (HCl(aq)36% w/w, Labsynth) and extracted 3 times with 20 mL dichloromethane (Labsynth). The organic solvent was removed under a N2(g) flux and the concentrate resuspended in 1 mL DMSO (Labsynth) and 2 mL methanol (Labsynth).
Dimethyl sulfoxide (dmso)
Manufactured by Labsynth
Sourced in Brazil
DMSO is a colorless, polar aprotic solvent that is widely used in various laboratory applications. Its primary function is to serve as a versatile solvent capable of dissolving a wide range of organic and inorganic compounds. DMSO has a high boiling point and is miscible with water and many other solvents.
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Lab products found in correlation
4 protocols using dimethyl sulfoxide (dmso)
1
Extraction of Metabolites from Malaria Parasite
2
Synthesis and Use of H2S Donors
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AP39 was synthesized in-house, as previously described [31 (link)]. All other drugs and reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA), except for apamin (Bio-Techne, Abingdon, UK). Stock solutions of the H2S donors AP39 and ADT-OH were prepared at 10 mM in 100% dimethylsulfoxide (DMSO; Labsynth, Diadema, Brazil), kept at −80 °C, and diluted to the final concentrations with Krebs-Henseleit solution just before use. All other compound solutions were freshly prepared just before use at concentrations 1000-fold higher than the final used concentrations. apamin, glibenclamide, ODQ, and sildenafil were firstly dissolved in 100% DMSO; L-NAME, TEA, and AOAA were dissolved in distilled water, and indomethacin was dissolved in 10% Na2CO3.
3
Amphotericin B Treatment of P. brasiliensis Infection in Larvae
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At 1 h after infection with P. brasiliensis, larvae were inoculated with amphotericin B (0.5 mg/kg) in the last right pro-leg. Amphotericin B stock solutions were prepared in dimethyl sulfoxide (DMSO) (Labsynth, Diadema, SP, Brazil) and diluted in PBS to a DMSO concentration of 5%, as described previously [14 (link)]. A group of uninfected larvae was treated with an antifungal agent alone to assess toxicity. The larvae were incubated at 37 °C for 7 days to prevent physical movement. Additionally, within 48 h of treatment with amphotericin B, haemolymph samples were collected through puncture of the larvae’s abdomen, and haemocyte density was established through microscopy using a Neubauer haemocytometer.
4
Starch Molecular Mass Distribution Analysis
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The molecular mass distribution profiles of the starch samples were determined by gel permeation chromatography (GPC) according to Song and Jane (2000) (link), with modifications. A GE XK 26/70 column (2.6 cm diameter and 70 cm high) packed with
Sepharose CL-2B gel (Sigma, Sweden) was used. 10 mL of dimethylsulfoxide (DMSO; 90%, Labsynth, Brazil) was added to 0.1 g of starch and heated in boiling water bath for 1 h, then remaining for 24 h at 25°C under constant stirring. An aliquot of 3 mL (30 mg of starch) was then mixed with 10 mL of absolute ethanol to precipitate the starch, being the suspension centrifuged for 30 min at 3000 g. The precipitated starch was dissolved in 9 mL of boiling distilled water and put in boiling water bath for 30 min. An aliquot of 4 mL was then eluted in the chromatographic column upwardly. A solution containing 25 mmol•L -1 of NaCl and 1 mmol•L -1 of NaOH was used as eluent at a rate of 60 mL•h -1 . Fractions of 4 mL were collected by a fraction collector (Gilson, model FC203B, Middleton, England) and analysed for total carbohydrate content at 490 nm by the phenol sulfuric method (DuBois, Gilles, Hamilton, Rebers, & Smith, 1956) (link) and blue value at 620 nm (Juliano, 1971) , using a microplate reader (Asys Expert plus, Biochron, England). A detector substance (glucose, in this case) was used to mark the end of the elution of the sample.
Sepharose CL-2B gel (Sigma, Sweden) was used. 10 mL of dimethylsulfoxide (DMSO; 90%, Labsynth, Brazil) was added to 0.1 g of starch and heated in boiling water bath for 1 h, then remaining for 24 h at 25°C under constant stirring. An aliquot of 3 mL (30 mg of starch) was then mixed with 10 mL of absolute ethanol to precipitate the starch, being the suspension centrifuged for 30 min at 3000 g. The precipitated starch was dissolved in 9 mL of boiling distilled water and put in boiling water bath for 30 min. An aliquot of 4 mL was then eluted in the chromatographic column upwardly. A solution containing 25 mmol•L -1 of NaCl and 1 mmol•L -1 of NaOH was used as eluent at a rate of 60 mL•h -1 . Fractions of 4 mL were collected by a fraction collector (Gilson, model FC203B, Middleton, England) and analysed for total carbohydrate content at 490 nm by the phenol sulfuric method (DuBois, Gilles, Hamilton, Rebers, & Smith, 1956) (link) and blue value at 620 nm (Juliano, 1971) , using a microplate reader (Asys Expert plus, Biochron, England). A detector substance (glucose, in this case) was used to mark the end of the elution of the sample.
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