The anesthetics ketamin and xylazin were obtained as chloridrate. Ketamin, sold as Dopalen 100 mg/mL, was obtained from Ceva Animal Health Ltd (São Paulo, Brazil), and xylazin (Coopazine 20mg/mL) came from Coopers (São Paulo, Brazil). Butylated hydroxytoluene (BHT) was purchased from Merck (Brazil); ascorbic acid (Citroplex), from Neo Química Laboratory (Goiás, Brazil); and 1,1-Diphenyl-2-picryl-hydrazyl (DPPH from Sigma-Aldrich (Brazil).
Dopalen
Manufactured by Ceva
Sourced in Brazil
Dopalen is a laboratory instrument used for the analysis of molecular weight distribution in polymer samples. It functions by employing the principles of dynamic light scattering to measure the hydrodynamic size of molecules in solution.
Automatically generated - may contain errors
Lab products found in correlation
Anasedan, by Ceva (9 mentions)
1 1 diphenyl 2 picrylhydrazyl dpph, by Merck Group (1 mentions)
Butylated hydroxytoluene, by Merck Group (1 mentions)
Proxymetacaine hydrochloride, by Alcon (1 mentions)
Calcium chloride dihydrate, by Merck Group (1 mentions)
Soy phosphatidylcholine, by Lipoid (1 mentions)
Neutral red, by Merck Group (1 mentions)
Middlebrook oadc, by Merck Group (1 mentions)
Oleylamine, by Merck Group (1 mentions)
Rifampicin, by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Potassium chloride (kcl), by Merck Group (1 mentions)
Croton oil, by Merck Group (1 mentions)
2 2 azino bis 3 ethylbenzothiazoline 6 sulfonic acid, by Merck Group (1 mentions)
Ultra fine 2, by BD (1 mentions)
Xylocaine 1, by Cristália (1 mentions)
Thiopentax, by Cristália (1 mentions)
12 protocols using dopalen
1
Anesthetic and Antioxidant Compounds Evaluation
2
Myocardial Infarction and Membrane Implantation
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The rats were anesthetized by intramuscular administration of ketamine (Dopalen®, Ceva Santé Animale, Paulínea, SP, Brazil) (50 mg/kg) and xylazine (Anasedan®, Ceva Santé Animale, Paulínea, SP, Brazil) (10 mg/kg) and were then subjected to mechanical ventilation. These procedures were followed by thoracotomy at the left, fifth intercostal space. To induce MI (D0), the left anterior descending (LAD) coronary artery was directly ligated just beyond the first diagonal branch. The rats were then maintained in cages and kept under controlled temperature and high oxygen pressure to facilitate postoperative recovery. Both normal respiratory activity and heart rate (350–450 beats/min) were carefully monitored before housing the animals under standard conditions until the next experimental step.
Seven days after surgery, the rats were subjected to another left thoracotomy (T2) for membrane implantation in the ventricular surface, with or without cocultured cells. Immediately after the LAD ligation, a bacterial membrane fragment was gently placed onto the left ventricle of group III animals, combined with cocultured cells in contact with the epicardial surface. The membrane fragment edges were ligated to the ventricle using a suture and placed without any artificial reinforcing effect. Finally, the sternum and skin incisions were sutured (Figure 2 ).
Seven days after surgery, the rats were subjected to another left thoracotomy (T2) for membrane implantation in the ventricular surface, with or without cocultured cells. Immediately after the LAD ligation, a bacterial membrane fragment was gently placed onto the left ventricle of group III animals, combined with cocultured cells in contact with the epicardial surface. The membrane fragment edges were ligated to the ventricle using a suture and placed without any artificial reinforcing effect. Finally, the sternum and skin incisions were sutured (
3
Intravitreal Toxicity of PaPn Peptides
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In order to investigate the intravitreal toxicity of the PaPn11 and PaPn13
peptides, the animals were anesthetized via intraperitoneal injection of 90mg/kg
ketamine (Dopalen; Ceva, Brazil) plus 10 mg/kg xylazine hydrochloride (Anasedan;
Ceva, Brazil). Sequentially, the right eyes were anesthetized using one drop of
0.5% (w/v) proxymetacaine hydrochloride (Anestalcon; Alcon, Brazil). For
intravitreal injections, the animals were divided into three main groups: PnPa11
(n = 20), PnPa13 (n = 20) and vehicle (saline) (n = 4). The groups that received
the synthetic peptide were subdivided according to the intravitreal
concentration of the peptides administrated (0.50; 1.25; 2.50; 3.75 and 5.00
µg/mL) (n = 4). The left eyes of all animals were kept intact.
A 30-gauge needle attached to a syringe was inserted ∼2 mm to the limbus. Besides
that, the needle was held in place for 30seconds to prevent it from escaping the
application site. The volume of intravitreal injection was set to 10 µL [28 (link)]. Each concentration was calculated
based on the dilution that occurs in the vitreous humor (for adult rats, the
vitreous volume is about 50 μL) [29 ].
peptides, the animals were anesthetized via intraperitoneal injection of 90mg/kg
ketamine (Dopalen; Ceva, Brazil) plus 10 mg/kg xylazine hydrochloride (Anasedan;
Ceva, Brazil). Sequentially, the right eyes were anesthetized using one drop of
0.5% (w/v) proxymetacaine hydrochloride (Anestalcon; Alcon, Brazil). For
intravitreal injections, the animals were divided into three main groups: PnPa11
(n = 20), PnPa13 (n = 20) and vehicle (saline) (n = 4). The groups that received
the synthetic peptide were subdivided according to the intravitreal
concentration of the peptides administrated (0.50; 1.25; 2.50; 3.75 and 5.00
µg/mL) (n = 4). The left eyes of all animals were kept intact.
A 30-gauge needle attached to a syringe was inserted ∼2 mm to the limbus. Besides
that, the needle was held in place for 30seconds to prevent it from escaping the
application site. The volume of intravitreal injection was set to 10 µL [28 (link)]. Each concentration was calculated
based on the dilution that occurs in the vitreous humor (for adult rats, the
vitreous volume is about 50 μL) [29 ].
4
Quantifying Nitrite Levels in Rat Plasma
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After the functional parameter measurements, all rats were deeply anesthetized by an overload of xylazine hydrochloride and ketamine hydrochloride (Anasedan®, 20 mg/kg and Dopalen®, 160 mg/kg, i.v., respectively, Ceva Sante Animale, Paulínea, SP, Brazil) and euthanized by decapitation. Blood samples were collected in heparinized vacutainers immediately after the euthanasia and centrifuged at 4,000 rpm for 5 min for analysis of nitrite concentration as previously published (Jacomini et al., 2017 (link)). In summary, proteins were quantified using automated biochemistry equipment (model A-15, Biosystems S/A, Barcelona, Spain) to normalize the calculation of nitrite concentration. Nitrites (NO2–), metabolites of NO, were determined in plasma using Griess reagent in which a chromophore with a strong absorbance at 540 nm is formed by the reaction of nitrite with a mixture of naphthyl ethylenediamine (0.1%) and sulfanilamide (1%). Samples were analyzed in duplicate, and plasma results are expressed as nmol/mg of protein.
5
Mice Husbandry and Experimental Procedures
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Male C57Bl/6 mice were obtained from the Center for Development of Animal Models for Medicine and Biology (CEDEME, Federal University of São Paulo). They were kept at the animal house of the Department of Bioscience in a temperature-controlled room (22°C) with a 12 h-12 h light-dark cycle (lights on 07.00-19.00 h) and free access to water and a standard diet with the following composition: 16.5% fat, 65.7% carbohydrate, 17.7% protein and a caloric density of 3.82 kcal g -1 . All mice were allowed to acclimate to the animal house for 2 weeks before commencement of any experiments. At the end of the experiment, mice were anaesthetized with a mixture of ketamine (Dopalen, 100 mg kg -1 , 1.0 ml kg -1 ; Ceva, Paulinia, São Paulo, Brazil) and xylazine (Anasedan, 10 mg kg -1 , 0.5 ml kg -1 ; Ceva) administered I.P., and all mice were killed by decapitation.
6
Mycobacterial Growth Inhibition Assay
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Rifampicin, purity ≥ 97% (Sigma Aldrich, St. Louis, MO, USA); Heparin, 5000 U.I./mL, (BlauFarmacêutica®, Cotia, Brazil); Dopalen, ketamine 10% injectable, veterinary use (Ceva, Paulínia, Brazil); Calmiun, xylazine 2% injectable, veterinary use (Agener União Saúde Animal, São Paulo, Brazil); Bacter Bacterial agar (Becton Dickinson, East Rutherford, EUA); Cetylalcoholethoxylated 20 OE and propoxylated 5 OP-Procetyl AWS (Croda, Campinas, Brazil); Calcium chloride dihydrate (Merck, St. Louis, CO, USA); Potassium chloride (Merck); Sodium Chloride (Merck); soy phosphatidylcholine (Lipoid GMBH, Geneva, Switzerland); culture medium-Dulbecco’s Modified Eagle Medium (Hyclone, Logan, UT, USA); Brook Middlebrook culture medium-7H11 (Difcot, St. Louis, CO, USA); Oleylamine (Sigma Aldrich, St. Louis, CO, USA); Middlebrook-OADC (Sigma Aldrich, St. Louis, MO, USA); Neutral Red (Sigma Aldrich, St. Louis, MO, USA).
7
Investigating Uvaol's Therapeutic Potential
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The following substances, purchased from Sigma Chemical Co. (St. Louis, MO, USA), were used: uvaol (Urs-12-ene-3,28-diol, ≥95% purity, PubChem CID: 92802; Figure 1 ), Tween-20, eosin, hematoxylin, Dulbecco’s Modified Eagle Medium (DMEM), Roswell Park Memorial Institute medium (RPMI-1640), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), phosphate-buffered saline (PBS) gentamicin, fetal bovine serum (FBS), trypsin, l -glutamine, 4′,6-diamidino-2′-phenylindole dihydrochloride (DAPI), SB203580 (p38-MAPK inhibitor), and (6-22)-amide (inhibitor of PKA). Xylazine (Anasedan®) and ketamine (Dopalen®) from Ceva (Paulínia, São Paulo, BRA), ethylenediaminetetraacetic acid (EDTA) and dimethyl sulfoxide (DMSO) from Synth (Diadema, SP, BRA). Antibodies against laminin, fibronectin, and collagen type I were purchased from Novotec (Bron, Lyon, France), while anti-goat IgG-fluorescein isothiocyanate (FITC) was from Santa Cruz Biotechnology (Dallas, TX, USA). Stock solution of uvaol was prepared in DMSO. The DMSO concentration applied to cells in culture never exceeded 0.1%. Neither the vehicle nor any of the compounds used in this study altered cell viability.
8
Spinal Cord Tissue Processing and Analysis
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After de end of the treatments, mice were anesthetized with Anasedan (xylazine, Ceva, Brazil, 10 mg/kg) and Dopalen (ketamine, Ceva, Brazil, 50 mg/kg) and subjected to transcardial perfusion with 0.9% saline in 0.1 M phosphate buffer (PBS), followed by fixative solution (4% paraformaldehyde in 0.1 M phosphate buffer—PB; pH 7,4). The lumbar spinal cords were removed, post-fixed in the same fixative solution for 12 h at 4 °C, washed with phosphate buffer (PB), and sequentially cryopreserved in 10, 20, and 30% PB-sucrose solution (12 h in each concentration).
Samples were individually frozen in n-hexane cooled in liquid nitrogen at −35°C. Transverse sections (12 μm thick) of the lumbar spinal cords were obtained in a cryostat (Microm HM525), transferred to gelatin-coated slides, dried at room temperature for 30 min, and stored at −20°C until use.
Neuronal survival analyses and immunofluorescence staining quantifications were performed in the ventrolateral and ventromedial region of the ventral horn in the spinal cord, where the motor neurons are located, as demonstrated inFigure 1 .
Samples were individually frozen in n-hexane cooled in liquid nitrogen at −35°C. Transverse sections (12 μm thick) of the lumbar spinal cords were obtained in a cryostat (Microm HM525), transferred to gelatin-coated slides, dried at room temperature for 30 min, and stored at −20°C until use.
Neuronal survival analyses and immunofluorescence staining quantifications were performed in the ventrolateral and ventromedial region of the ventral horn in the spinal cord, where the motor neurons are located, as demonstrated in
9
Sesamol-Based Topical Formulation Evaluation
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Sesamol (CAS number: 533-31-1), sorbitan monooleate (Span® 80), 1-1-diphenyl-2-picrylhydrazyl (DPPH) radical, 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and croton oil were obtained from Sigma Aldrich (São Paulo, SP, Brazil). The ethylcellulose polymer (Ethocel®TM Standard 10 Premium Ethylcellulose) and guar gum were donated by Colorcon (Cotia, SP, Brazil) and CP Kelco (Limeira, RS, Brazil), respectively. The polysorbate 80 (Tween® 80) and medium-chain triglycerides (MCTs) were provided by Delaware (Porto Alegre, RS, Brazil). Dexamethasone acetate (1 mg/g; Teuto®) was purchased at a commercial drugstore. Ceva (Paulínia, SP, Brazil) was the supplier of the anesthetics ketamine (Dopalen®) and xylazine (Anasedan®). Vetec (Rio de Janeiro, RJ, Brazil) was the supplier of the formaldehyde, ethanol, sodium acetate and acetic acid. Merck (Darmstadt, Germany) was the supplier of hematoxylin–eosin and paraffin. Acetone was acquired from CRQ (São Paulo, SP, Brazil). The high-performance liquid chromatography grade methanol was acquired from LiChrosolv (São Paulo, SP, Brazil). All other reagents and solvents were of analytical grade and used as received.
10
Curcumin's Renoprotective Effects in CKD and IRI
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In the study, 20 male Wistar rats weighing 250-300 g were used, supplied by the Institute of Biomedical Sciences of the University of São Paulo (ICB-USP) and maintained with free access to water and feed, under thermal conditions with alternating day and night cycles. procedureS Induction of CKD -The animals were anesthetized with 10 mg/kg of xylazine (Anasedan, Vetbrands) and 90 mg/kg of ketamine (Dopalen, Ceva) intraperitoneal (IP) and submitted to the surgical technique of 5/6 renal ablation, which consists of laparotomy followed by right nephrectomy; surgical clamping of two branches of the left renal artery; and suture of the abdominal incision (10) (link) . Curcumin administration: curcumin was administered for 10 days, from the 17th day of the experiment, at a 30 mg/kg/day dose, orally (P.O.), diluted in 0.5% in volume carboxymethylcellulose solution 0.5 mL/100 g (9) (link) . Induction of acute renal injury by IRI -The animals were anesthetized, as described, and subjected to laparotomy for unilateral clamping of the renal pedicles for 30 minutes, with non-traumatic vascular clamps. The animals were then evaluated for anesthetic recovery and received analgesia in the postoperative period (Tramadol: 5 mg/kg, IP, 8/8 h, until the third postoperative day) (Figure 1) (7) (link) .
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