Thiopental

Manufactured by Abbott

Sourced in United States

Thiopental is a barbiturate drug used as a general anesthetic. It is a fast-acting, short-duration anesthetic agent that can be administered intravenously. Thiopental acts by enhancing the effects of gamma-aminobutyric acid (GABA), the primary inhibitory neurotransmitter in the central nervous system.

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Lab products found in correlation

Stemi dv4 stereo microscope, by Zeiss (1 mentions) Quad bridge amp, by ADInstruments (1 mentions) Powerlab, by ADInstruments (1 mentions)

8 protocols using thiopental

Kidney Histopathology After Tumor Implantation

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At the end of the experiment, all rats were weighed and euthanized with thiopental (40 mg/kg of body weight; Abbott Laboratories, Chicago, IL, United States of America), intraperitoneally administered. The Walker-256 solid tumor was removed and weighed. A laparotomy was performed to collect the kidneys, which were washed in PBS, weighed, submitted to fixation, and destined for processing.
After 48 hours of fixation in 10% buffered formalin solution, the kidneys were coronally sectioned, and the specimens were destined for histological routine with dehydration in an increasing series of alcohols (80%, 90%, absolute I, II and III), clearing in xylene (three times) and embedded with paraffin. Slides with 3-μm thick sections were obtained and destined periodic acid-Schiff (PAS), Masson’s trichrome, and immunohistochemical staining28 (link).

Lima K.R., Lopes M.L., de Souza S.R., Fracaro L., da Purificação N.R., Lima M.F., Lins L.A., Lacchini S., de Araújo A.A., de Araújo RF J.ú.n.i.o.r., Perles J.V., Zanoni J.N, & Clebis N.K. (2023). L-glutamine supplementation reduced morphological damage in the renal glomerulus of rats with Walker-256 tumor. Acta Cirúrgica Brasileira, 38, e383923.

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Histological Analysis of Jejunal Innervation

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After a period of 120 days, the animals were weighed and euthanized under anesthesia with thiopental (40 mg/kg body weight, i.p.; Abbott Laboratories, Chicago, IL, USA). The blood was collected by using cardiac puncture for the blood glucose measurements (20 (link)). After celiotomy, the samples were collected from all the animals, and then washed with phosphate-buffered saline (PBS, 0.1M, pH 7.4), gently inflated with Zamboni’s fixative solution to fill the space previously occupied by the stool, thus avoiding the jejunal stretching. After that, the jejunums were maintained for 18 h in the same solution at 4°C. After fixation, the samples were carefully opened along the mesenteric border and successively rinsed with 80% ethanol in order to completely remove the fixative excess. Then, the dehydrations were sequentially performed with ethanol concentration of 95 and 100%, followed by clarification in xylene. After that, the rehydration was performed by using ethanol at decreasing concentrations (100, 90, 80, and 50%). The tissues were stored at 4°C in PBS with 0.08% sodium azide.
The samples were microdissected under a stereomicroscope in order to obtain the whole mounts of muscularis tunica by the removal of the mucosal and submucosal layers. The tissues were double stained by immunohistochemistry to label HuC/D and S-100 proteins (24 (link), 25 (link)).

de Souza S.R., de Miranda Neto M.H., Martins Perles J.V., Vieira Frez F.C., Zignani I., Ramalho F.V., Hermes-Uliana C., Bossolani G.D, & Zanoni J.N. (2017). Antioxidant Effects of the Quercetin in the Jejunal Myenteric Innervation of Diabetic Rats. Frontiers in Medicine, 4, 8.

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Assessing Tumor-Induced Cachexia in Rats

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At the end of the experimental period of 14 days, the animals were weighed and received an injection of vincristine sulfate (0.5 mg.kg⁻¹ body weight; Tecnocris^™, Eurofarma/Zodiac Laboratories, São Paulo, SP, Brazil) two hours before euthanasia. The administration of this microtubule assembly inhibitor improves immunohistochemistry. Rats were anesthetized with thiopental (40 mg.kg⁻¹ body weight i.p.; Abbott Laboratories, Chicago, IL, USA), tissues were harvested and the animal was killed via thoracotomy. The tumor mass was removed from the animals and then weighed to calculate the percent (%) loss of body mass index (cachexia index) according to the equation:^{39 (link)}Where: ibm = initial body mass of the tumor-bearing animal; fbm = final body mass of the tumor-bearing animal; mt = mass of the tumor; gmc = the mean mass gain of the control group (C). Jejunal tissues were collected, washed in PBS (0.1M, pH 7.4).

Fracaro L., Frez F.C., Silva B.C., Vicentini G.E., de Souza S.R., Martins H.A., Linden D.R., Guarnier F.A, & Zanoni J.N. (2015). Walker 256 tumor-bearing rats demonstrate altered interstitial cells of Cajal Effects on ICC in the Walker 256 tumor model. Neurogastroenterology and motility : the official journal of the European Gastrointestinal Motility Society, 28(1), 101-115.

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Intestinal and Adipose Tissue Analysis

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After fasting for 15 h, the animals received 0.5 mg/kg of vincristine sulfate solution 2 h prior to euthanasia (Tecnocris, Eurofarma, São Paulo, SP, Brazil) intraperitoneally to block mitosis in the intestinal epithelium. The animals were weighed and intraperitoneally anesthetized with 80 mg/kg Thiopental (Abbott Laboratories, Chicago, IL, USA) and underwent laparotomy to collect the small intestine and periepididymal, retroperitoneal, and mesenteric fat deposits. The fat deposits were weighed, and the lengths (from the pylori to the ileo-cecal junction) of the small intestine were measured. Samples of the duodenum, jejunum, and ileum were opened at the mesenteric border immediately after collection and the intestinal circumference was measured. From the 10 animals in each group, five underwent immunohistochemical procedures, and the other five underwent histological procedures.

Soares A., Beraldi E.J., Ferreira P.E., Bazotte R.B, & Buttow N.C. (2015). Intestinal and neuronal myenteric adaptations in the small intestine induced by a high-fat diet in mice. BMC Gastroenterology, 15, 3.

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L-Glutamine Supplementation in Walker 256 Tumor-Bearing Rats

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This study was approved by the Ethics Committee on Animal Use of the Universidade Estadual de Maringá (protocol nº 099/2012). Forty sexually mature male Wistar rats (Rattus norvegicus) at the age of 55 days were obtained from the Central Biotery of the State University of Maringá and kept under controlled illumination (12h light / 12h dark) and temperature (22±2ºC). The animals were housed in polypropylene cages and supplied with water and food ad libitum. The animals were randomized into four experimental groups (n=10): control without L-glutamine (C); control supplemented with L-glutamine (CG); inoculated with Walker 256 tumor cells (WT) and inoculated with Walker 256 tumor cells and supplemented with L-glutamine (WTG). The animals received daily treatment for 14 days. At the end of the experimental period, animals were fasted for 12 h before being weighed and euthanized under intraperitoneal anesthesia (40 mg/kg body weight thiopental – Abbott Laboratories, Chicago, IL, USA).

Rocha N.R., Braz J.K., de Souza S.R., Fracaro L., de Melo F.C., Zanoni J.N., Clebis N.K., Morais D.B, & de Moura C.E. (2021). Testicular morphometry of rats with Walker 256 tumor supplemented with L-glutamine. Animal Reproduction, 18(2), e20200051.

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Vincristine Effects on Rat Metabolism

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After 60 days of experiment, the rats were weighed and vincristine (0.5 mg/kg -Eurofarma Laboratories Ltda, São Paulo, SP, Brazil) was intravenously administered 2 h prior to their euthanasia. After that, the rats were anesthetized with thiopental (40 mg/kg -Abbott Laboratories, Chicago, IL, USA) intraperitoneally. Blood samples were obtained by cardiac puncture to measure the blood glucose levels using glucose-dyeoxidoreductase technique (Bioclin, Belo Horizonte, MG, Brazil) (Pereira et al., 2011) (link). After celiotomy, the ileal segments were obtained for the immunohistochemistry and oxidative stress techniques.

Sehaber-Sierakowski C.C., Vieira-Frez F.C., Hermes-Uliana C., Martins H.A., Bossolani G.D.P., Lima M.M., Blegniski F.P., Guarnier F.A., Baracat M.M., Perles J.V.C.M, & Zanoni J.N. (2021). Protective effects of quercetin-loaded microcapsules on the enteric nervous system of diabetic rats. Autonomic neuroscience : basic & clinical, 230.

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Evaluation of Enteric Innervation in Ileum

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After 60 days of experiment, animals were anesthetized with Thiopental (40 mg kg/kg body weight; Abbott Laboratories, Chicago, IL, USA) and the ileum was collected. After tissue collection, intestinal samples were rinsed in phosphate-buffered saline (PBS 0.1M, pH 7.4; 10 mmol/L Na 2 HPO 4 and 150 mmol/L NaCl), followed by incubation in Zamboni's fixative for 18 h at 4ºC and then, washed in 80% alcohol. Whole-mounts were sequentially dehydrated in 95% and 100% alcohol, diaphanized in xylene and then successively rehydrated in 100%, 90%, 80% and 50% alcohol. Afterwards, ileal samples were cut along the mesenteric border and rinsed in PBS for 30 min and then, stored in PBS with 0.08% sodium azide at 4ºC. Subsequently, intestinal whole-mounts were dissected under a Stemi DV4 stereomicroscope (Zeiss, Jena, Germany) containing outer muscle layers and the myenteric plexus. Ten ileal segments were dissected and processed for the subsequent immunohistochemistry technique to the nNOS, VIP and CGRP.

Zanoni J.N, & Piovezana Bossolani G.D. (2019). DOES THE RHEUMATOID ARTHRITIS AFFECT THE ENTERIC NERVOUS SYSTEM?. Arquivos de gastroenterologia, 56(2).

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Anesthetized Rat Hemodynamic Monitoring

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Male Sprague-Dawley rats (100-130 g) were randomly allocated to treatment groups (Table S1; n = 4-7). The study protocol was approved by the Flinders University Animal Welfare Committee.
Rats were anaesthetised via continuous intravenous infusion of thiopental (60 mg/kg/h; Abbott) and paralysed with a bolus injection of pancuronium bromide (1 mg/kg i.v.; Astra Zeneca, UK) maintained by continuous infusion (0.2 mg/ kg/h i.v.). Mean arterial pressure was continuously monitored through cannulation of the left femoral artery using disposable blood pressure transducers connected to a Pow-erLab and Quad Bridge Amp (AD Instruments, Australia).

Elder A.S.F., Bersten A.D., Saccone G.T.P., Bonder C.S, & Dixon D.L. (2019). Prevention and Amelioration of Rodent Ventilation-Induced Lung Injury with Either Prophylactic or Therapeutic feG Administration. Lung, 197(5).

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