Carotid artery reendothelialization was evaluated after perivascular electric injury in 12‐ to 16‐week‐old male mice as previously described.32 (link)–33 (link) Mice were anesthetized via intraperitoneal administration of avertin, and the left common carotid artery was exposed through an anterior incision of the neck. Electric current of 4 W was applied through 2‐mm forceps (2 W/mm) for 3 sec in microbipolar mode (Force 2 Electrosurgical Generator; Valleylab). The mice were allowed to recover, and analgesia was provided via intraperitoneal administration of buprenorphine immediately after surgery and 24 hours later. At 72 hours after injury, animals were injected intravenously with 5% Evans blue dye (Sigma‐Aldrich), the carotid arteries were harvested, and the area of denudation (which incorporates the dye) was quantified in a blinded manner through image analysis by using Scion Image (free software from National Institutes of Health). NHIgG or aPLs (100 μg per mouse) were injected intraperitoneally 24 hours before injury, on the day of injury, and 24 hours after injury. For evaluation of the effect of molsidomine on reendothelialization, mice were injected intraperitoneally with molsidomine (Sigma‐Aldrich, 5 mg/kg per day) or vehicle (saline) daily starting 72 hours before injury and ending 48 hours after injury.
Molsidomine
Manufactured by Merck Group
Sourced in United States
Molsidomine is a synthetic organic compound that functions as a prodrug, which is converted into an active metabolite that can stimulate the release of nitric oxide. It is commonly used in laboratory settings for research and experimentation purposes.
Automatically generated - may contain errors
Lab products found in correlation
Clozapine, by Merck Group (3 mentions)
Risperidone, by Merck Group (2 mentions)
Evans blue dye, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Ketamine hydrochloride, by Merck Group (1 mentions)
L name, by Merck Group (1 mentions)
96 well polystyrene plate, by Thermo Fisher Scientific (1 mentions)
Mahma nonoate, by Merck Group (1 mentions)
8 protocols using molsidomine
1
Carotid Artery Reendothelialization after Perivascular Electric Injury
2
Preparation of Psychoactive Drug Solutions
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Apomorphine hydrochloride was dissolved in saline (NaCl 0.9%) that contained 0.1% ascorbic acid to prevent oxidation. Molsidomine (Sigma, St. Louis, MO, USA) was dissolved in saline (NaCl 0.9%). Clozapine (Sigma, St. Louis, MO, USA) and risperidone (Sigma, St. Louis, MO, USA) were dissolved in a minimum volume of acetic acid, made up to volume with saline, and pH adjusted to 7 with 0.1 M NaOH. All drug solutions were freshly prepared on the day of testing and were administered intraperitoneally (i.p.) in a volume of 1 mL/kg. For all studies, control rats received isovolumetric quantities of the specific vehicle solutions. The chemical structures of Molsidomine, Clozapine, and risperidone are illustrated in Figure 5 .
3
Screening for Biofilm-Disrupting NO Donors
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The following criteria were used to select candidate nitric oxide donors: (1) low/moderate toxicity; (2) no more than 0.1 % of probable, possible or confirmed human carcinogenicity according to the International Agency for Research on Cancer (IARC); (3) low/moderate cost; (4) commercial availability.
Nitric oxide donors used in this study: S-nitroso-N-acetyl-d ,l -penicillamine (SNAP) (Cayman Chemicals, Ann Arbor, MN, USA), 3-(aminopropyl)-1-hydroxy-3-isopropyl-2-oxo-1-triazene (Noc-5), 2-(acetyloxy)benzoic acid 4-(nitroxymethyl)phenyl ester (NO-aspirin), 6-(2-hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-hexanamine (MAHMA NONOate), and molsidomine (all from Sigma-Aldrich, St. Louis, MO, USA). For each compound, 1 mM stock solutions were prepared in phosphate-buffered saline, pH 7.3 (PBS, Fisher, Waltham, MA, USA) and aliquots were stored at −80 °C. For the assays, serial dilutions were always prepared fresh in PBS just before the experiments and used within 5 min of their preparation. Biofilm dispersion potential of the molecules was tested on polystyrene and polypropylene 96 well-plates (Fisher, Waltham, MA, USA).
Nitric oxide donors used in this study: S-nitroso-N-acetyl-
4
Oxidative Stress Induction in Mice
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Oxidative stress was induced with administration of molsidomine (SIN-10) (Sigma Aldrich Canada). Mice aged 3 or 18 mo were given a single intra-peritoneal injection of 30 mg/kg of body weight 2 h prior to being euthanized [37 (link)]. SIN-10 treatment time was chosen based on previous studies that indicated that serum levels of the SIN-10 metabolite SIN-1 peaked at 2 h [38 (link)], and experimental trials in which the treatment time of 2 h was found to induce a peak increase in ROS in pachytene spermatocytes from young WT animals (Supplemental Figure S1A; Supplemental Data are available online at www.biolreprod.org ). The SIN-10 dose was based on information in the literature [37 (link), 38 (link)] and preliminary dose-response trials using aged animals to measure ROS response to the oxidative insult (Supplemental Figure S1B). In control animals, SIN-10 was replaced with saline injections. After each treatment, mice were euthanized using carbon dioxide followed by cervical dislocation. Tissues were removed, weighed, flash frozen, and stored at −80°C for later analyses. Five to seven animals were used for each of the four groups of animals (young and old serving as either control or SIN-10 treated) for both the Sod−/− and the Cat−/−studies.
5
Intraperitoneally Administered Drug Preparation
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Ketamine hydrochloride and molsidomine (Sigma, St. Louis, MO, USA) were dissolved in saline (NaCl 0.9%). Clozapine (Sigma, St. Louis, MO, USA) was dissolved in a minimum volume of acetic acid, made up to a volume with saline and pH adjusted to 7 with 0.1 M NaOH. All drug solutions were freshly prepared on the day of testing and were administered intraperitoneally (i.p.) in a volume of 1 mL/kg. For all studies, control animals received isovolumetric amounts of the specific vehicle solutions.
6
Preparation of Psychoactive Drug Solutions
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Apomorphine hydrochloride was dissolved in saline (NaCl 0.9%) that contained 0.1% ascorbic acid to prevent oxidation. Molsidomine (Sigma, St. Louis, MO, USA) was dissolved in saline (NaCl 0.9%). Clozapine (Sigma, St. Louis, MO, USA) and risperidone (Sigma, St. Louis, MO, USA) were dissolved in a minimum volume of acetic acid, made up to volume with saline, and pH adjusted to 7 with 0.1 M NaOH. All drug solutions were freshly prepared on the day of testing and were administered intraperitoneally (i.p.) in a volume of 1 mL/kg. For all studies, control rats received isovolumetric quantities of the specific vehicle solutions. The chemical structures of Molsidomine, Clozapine, and risperidone are illustrated in Figure 5 .
7
Pharmacological Modulation of L-NAME, Molsidomine, and CI-994
8
Preparation and Evaluation of Biofilm Dispersion Compounds
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MAHMA NONOate and molsidomine were purchased from Sigma-Aldrich (St. Luois, MO, USA). For each compound, 1 mmol L−1 stock solutions were prepared in phosphate-buffered saline (PBS), pH 7.3 (PBS, Fisher, Waltham, MA, USA) and aliquots were stored at −80°C. For the assays, serial dilutions were always prepared fresh in ice-cold PBS just before the experiments and used within 5 min of their preparation. The biofilm dispersion potential was tested on polystyrene 96-well plates (Fisher, Waltham, MA, USA).
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