All animals were euthanized within 6 weeks post-surgery by an intraperitoneal overdose (150 mg/kg body weight) of the anesthetic: xylazine–ketamine and thiopental (Thiopentax™, Cristália, Itapira, Brazil). After euthanasia of the animals, the skull was dissected and removed for macroscopic, radiographic, and tomographic evaluations. Next, specimens of interest were removed, fixed in 10% buffered formaldehyde, and processed for microscopic analysis.
Thiopentax
Manufactured by Cristália
Sourced in Brazil
Thiopentax is a laboratory equipment product manufactured by Cristália. It is a device used for the administration of thiopental sodium, a fast-acting barbiturate used for the induction of general anesthesia.
Automatically generated - may contain errors
Lab products found in correlation
Compaz, by Cristália (2 mentions)
Vecuron, by Cristália (2 mentions)
Fenchone, by Merck Group (1 mentions)
Cyclophosphamide, by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Tween 80, by Merck Group (1 mentions)
5 fluorouracil 5 fu, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Mallinckrodt (1 mentions)
Xylocaine 1, by Cristália (1 mentions)
Dopalen, by Ceva (1 mentions)
Anasedan, by Ceva (1 mentions)
Humulin r, by Eli Lilly (1 mentions)
Accu chek active, by Roche (1 mentions)
12 protocols using thiopentax
1
Euthanasia and Tissue Harvesting
2
Pharmacological Evaluation of Anticancer Agents
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Propidium iodide (P4170 Sigma-Aldrich), 5-Fluorouracil (5-FU) (F6627 Sigma-Aldrich), Triton X-100 (93,443 Sigma-Aldrich), Tween 80 (P4780 Sigma-Aldrich), and cyclophosphamide (C7397 Sigma-Aldrich), Dimethylsulfoxide (DMSO) (67–68-5 Mallinckrodt Chemicals®), Sodium thiopental (Thiopentax®) was purchased from Cristália (Itapira, SP, Brazil), and heparin (Parinex®) from Hipolabor (Sabará, MG, Brazil). Kits for biochemical and hematological analysis were purchased from LABTEST® (ALT/GPT Liquiform ref.: 108; ALT/GPT Liquiform ref.:1008; Creatinina ref.: 35; Uréia CE ref.: 27) (Lagoa Santa, MG, Brazil). (+)-Fenchone (analytical standard) (46,208 Sigma-Aldrich).
3
Gliclazide Ameliorates Experimental Periodontitis
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Stock solution of gliclazide (GLI) was obtained by dissolving 30 mg gliclazide (Servier, Rio de Janeiro, RJ, Brazil) in distilled water. Distilled water served as the vehicle in the NL and L groups. GLI or vehicle was administered by oral gavage (1 mL per rat) 1 h before ligature placement (induction of experimental PD), and once daily thereafter for 10 days. The animals were assigned randomly to the following five groups (n=10 each): NL, L, L with 1 mg/kg GLI, L with 5 mg/kg GLI, and L with 10 mg/kg GLI. Drug doses were selected based on those used in humans and in in vivo studies examining the effect of gliclazide in rats.20 The animals were euthanized 11 days after initial treatment with an injection of 80 mg/kg thiopental (0.5 g Thiopentax; Cristália, São Paulo, SP, Brazil). The maxillae were fixed in 10% buffered formalin for histopathological, immunohistochemical (IHC), and immunofluorescent morphological analyses. Rat maxillae were fixed in 10% buffered formalin for 24 h and stored in 70% alcohol for micro-computed tomography (micro-CT) analysis. Gingival tissues were frozen at -80°C for myeloperoxidase, malondialdehyde, glutathione, cytokine, and quantitative reverse transcription polymerase chain reaction (qRT-PCR) analyses.
4
Animal Euthanasia Procedure Protocol
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All procedures were conducted according to the guidelines for animal research of the Brazilian College of Animal Experimentation (Colégio Brasileiro de Experimentação Animal) and the National Council of Animal Experimentation Control (Conselho Nacional de Controle de Experimentação Animal). The IGM—FIOCRUZ Institutional Review Board for Animal Experimentation approved all procedures (CEUA—Instituto Gonçalo Muniz—IGM/FIOCRUZ—protocol number: 020/2011). At the end of the study all animals were euthanized. The dogs were sedated with acepromazine (0.1 mg/kg, iv, Acepram 1%, Vetnil, Brazil) and sodium thiopental (15 mg/kg, iv, Thiopentax 1 g, Cristália Brazil) and euthanized using a saturated solution of potassium chloride (2 mL/kg, iv).
5
Adipose Tissue Harvesting and Anesthesia in Rabbits
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A qualified veterinarian assisted with the anesthetic procedure. Prior to surgical incision for autologous adipose tissue collection, the rabbits were premedicated intramuscularly with a combination of ketamine 10 mg/kg (Dopalen®, 100 mg/mL, CEVA, Paulínia, SP, Brazil), xylazine hydrochloride 3 mg/kg (Anasedan® 2%, 20 mg/mL, CEVA, Paulínia, SP, Brazil), and acepromazine 0.1 mg/kg (Apromazin® 0.2%, 200 mg/mL, Barueri, SP, Brazil). Additionally, Lidocaine 7 mg/kg (Xylocaine 1% 20 mg/mL, Cristália, Butantã, SP, Brazil), a topical anesthetic, was administered at the projected site of the surgical incision.
During laparotomy, the rabbits were maintained under general inhalation anesthesia with 3–4% isoflurane (Isoforine®, Cristália, Itapira, SP, Brazil) diluted with 100% oxygen. An intravenous 0.9% saline solution was administered for hydration. Upon laparotomy completion, the rabbits were observed for 21 days and then euthanized with a thiopental lethal overdose (Thiopentax, 120 mg/kg, Cristália, Itapira, SP, Brazil).
During laparotomy, the rabbits were maintained under general inhalation anesthesia with 3–4% isoflurane (Isoforine®, Cristália, Itapira, SP, Brazil) diluted with 100% oxygen. An intravenous 0.9% saline solution was administered for hydration. Upon laparotomy completion, the rabbits were observed for 21 days and then euthanized with a thiopental lethal overdose (Thiopentax, 120 mg/kg, Cristália, Itapira, SP, Brazil).
6
Lung Mechanics Analysis in Anesthetized Rats
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One week after the last administration of saline or ghrelin (Fig. 1b ), animals were pre-medicated with diazepam [(10 mg kg-1, i.p.) Compaz®, Cristália, Itapira, SP, Brazil], anaesthetised with thiopental sodium [(20 mg kg-1 i.p.) Thiopentax®, Cristália, Itapira, SP, Brazil], tracheotomised, paralysed with Vecuronium bromide 0.005 mg kg-1 i.v. (Vecuron®, Cristália, Itapira, SP, Brazil), and ventilated with a constant flow ventilator (Samay VR15; Universidad de la Republica, Montevideo, Uruguay) set as follows: respiratory rate 100 breaths/min, tidal volume (VT) 0.2 mL, and fraction of inspired oxygen (FiO2) 0.21. The anterior chest wall was surgically removed and a positive end-expiratory pressure (PEEP) of 2 cmH2O applied. Airflow and tracheal pressure (Ptr) were measured. In an open chest preparation, Ptr reflects transpulmonary pressure (PL). Lung mechanics were analysed by the end-inflation occlusion method [25 (link), 26 (link)]. Static lung elastance (Est,L) was determined by dividing lung elastic recoil pressure (Pel) by VT. Est,L was measured 10 times in each animal. All data were analysed using ANADAT software (RHT-InfoData, Inc., Montreal, Quebec, Canada). All experiments lasted less than 15 min.
7
Insulin Sensitivity Evaluation in Rats
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KITT was performed at ZT2. To assess insulin sensitivity, rats were anesthetized with thiopental sodium (Thiopentax®, Cristália) (40 mg/kg of animal weight), followed by an injection of regular insulin solution (750 mU/mL per kg of body weight) (Humulin R; Eli Lilly, Paris, France) in the penile vein. Blood samples from the tail were collected at 0 (baseline), 4, 8, 12, and 16 min after the insulin injection and measured using a glucometer (Accu-Chek Active, Roche Diagnostics Brazil Ltda.). The fractional disappearance for plasma glucose (KITT) was calculated by the formula 0.693/t½ (where t½ is the half-life of plasma glucose calculated from the slope of the curve obtained during the linear decay of plasma glucose).41 (link),44 (link)
8
Kidney Function and Oxidative Stress
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At the end of the protocol (85 days), animals were placed in individual metabolic cages for 24-hour urine collection to evaluate renal function and oxidative metabolites. After that, animals were anesthetized by intraperitoneal (i.p.) route with Thiopentax® (Cristália, Brazil) (sodium thiopental: 40-50 mg/kg) for blood collection by means of a puncture in the abdominal aorta and later evaluation of renal function.
The right kidney was removed and prepared for histological sections stained with hematoxylin-eosin. Another aliquot was immediately cooled and stored at -80ºC for later quantification of antioxidant enzymes. At the end of the study, animals were euthanized using the ethical guidelines for handling of experimental animals mentioned above.
The right kidney was removed and prepared for histological sections stained with hematoxylin-eosin. Another aliquot was immediately cooled and stored at -80ºC for later quantification of antioxidant enzymes. At the end of the study, animals were euthanized using the ethical guidelines for handling of experimental animals mentioned above.
9
Calvaria Extraction and Microtomography Analysis
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After 14 and 42 days, five animals in each group were subjected to excessive doses of anesthetic, 2.5% sodium thiopental (Thiopentax®, Cristália, Produtos Químicos Farmacêuticos, São Paulo, Brazil) intraperitoneally, and an intramuscular injection of 1% lidocaine hydrochloride (Blau® Farmacêutica SA, São Paulo, Brazil).
The calvaria of each animal were removed preserving the supraperiosteal soft tissues and fixed in a 10% buffered formalin solution and later were destined for computerized microtomography analysis.
The calvaria of each animal were removed preserving the supraperiosteal soft tissues and fixed in a 10% buffered formalin solution and later were destined for computerized microtomography analysis.
10
Characterizing Lung Mechanics in Rodents
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One week after the last instillation, the animals were premedicated with diazepam 10 mg/kg i.p. (Compaz®, Cristália, Itapira, SP, Brazil) anesthetized with thiopental sodium 20 mg/kg i.p. (Thiopentax®, Cristália, Itapira, SP, Brazil), tracheotomized, paralyzed with Vecuronium bromide 0.005 mg/kg i.v. (Vecuron®, Cristália, Itapira, SP, Brazil), and ventilated with a constant flow ventilator (Samay VR15; Universidad de la Republica, Montevideo, Uruguay) using the following settings: rate 100 breaths/min, tidal volume (VT) 0.2 mL, and fraction of inspired oxygen (FiO2) 0.21. The anterior chest wall was surgically removed and a positive end-expiratory pressure of 2 cm H2O applied. Airflow and tracheal pressure (Ptr) were measured. Lung mechanics were analyzed by the end-inflation occlusion method. In an open chest preparation, Ptr reflects transpulmonary pressure (PL). Static lung elastance (Est,L) was determined by dividing lung elastic recoil pressure (Pel) by VT. Est,L was measured 10 times in each animal. All data were analyzed using ANADAT software (RHT-InfoData, Inc., Montreal, Quebec, Canada). All experiments lasted < 15 min.
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