When canine experiments were conducted, an initial intramuscular injection of atropine sulfate (Mitsubishi Tanabe Pharma, Osaka, Japan) was administered at 0.04 mg/kg as premedication, followed by an intramuscular injection of a three-mixture solution to induce anesthesia. This mixture included 0.2 mg/kg of butorphanol tartrate (Meiji Seika Pharma, Tokyo, Japan), 80 µg/kg of medetomidine hydrochloride (Nippon Zenyaku Kogyo, Fukushima, Japan), and 0.1 mg/kg of midazolam (Astellas Pharma, Tokyo, Japan). Under local anesthesia with lidocaine (Dentsply Sirona, Tokyo, Japan), upper canine teeth were extracted from 9-month-old female dogs (n = 3) (Kitayama Labes, Ina, Japan). DPSCs were isolated and expanded in T-flasks at 3% oxygen concentration as described previously [21 (link)] with a slight modification of not rotating the culture at the third passage. DPSCs were cryopreserved at 1 × 106 cells/mL at the 4th passage for allogeneic transplantation.
Atropine sulfate
Manufactured by Mitsubishi
Sourced in Japan
Atropine sulfate is a chemical compound used in various laboratory applications. It is a salt of the organic compound atropine, which is derived from the Atropa belladonna plant. Atropine sulfate is commonly used as a reference standard in analytical procedures and as a component in certain laboratory reagents.
Automatically generated - may contain errors
Lab products found in correlation
Midazolam, by Astellas Pharma (3 mentions)
Butorphanol tartrate, by Meiji Seika Pharma (2 mentions)
Medetomidine hydrochloride, by Nippon Zenyaku Kogyo (2 mentions)
Lidocaine, by Dentsply (1 mentions)
Dormicum, by Astellas Pharma (1 mentions)
Isoflo, by Zoetis (1 mentions)
Cerenia, by Zoetis (1 mentions)
Pentobarbital sodium, by Kyoritsu Seiyaku (1 mentions)
Xylazine, by Bayer (1 mentions)
Sevoflurane, by Pfizer (1 mentions)
Ketamine, by Daiichi Sankyo (1 mentions)
Fentanyl, by Daiichi Sankyo (1 mentions)
7 protocols using atropine sulfate
1
Canine Dental Stem Cell Isolation
2
Anesthetic Protocol for Animal Research
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Each animal received a pre-anesthetic dose of atropine sulfate (0.05 mg/kg,
subcutaneously; Mitsubishi Tanabe Pharma Co., Osaka, Japan) and, after 10 min of
pre-oxygenation, anesthesia was induced with intravenous midazolam (0.02 mg/kg; Astellas
Pharma Inc., Tokyo, Japan) and propofol (4–8 mg/kg; Fresenius Kabi Japan Co., Tokyo,
Japan), followed by endotracheal intubation. Anesthesia was maintained with sevoflurane
(2.5–3.0%; Mylan Pharma Co., Tokyo, Japan) in oxygen.
subcutaneously; Mitsubishi Tanabe Pharma Co., Osaka, Japan) and, after 10 min of
pre-oxygenation, anesthesia was induced with intravenous midazolam (0.02 mg/kg; Astellas
Pharma Inc., Tokyo, Japan) and propofol (4–8 mg/kg; Fresenius Kabi Japan Co., Tokyo,
Japan), followed by endotracheal intubation. Anesthesia was maintained with sevoflurane
(2.5–3.0%; Mylan Pharma Co., Tokyo, Japan) in oxygen.
3
Canine Adipose-Derived Mesenchymal Stem Cell Isolation
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All animal experimental protocols were approved by the Institutional Animal Care and Use Committee of the Osaka University Graduate School of Dentistry. Canine ADMPCs were isolated from greater omentum tissue. When harvesting this tissue, an initial intramuscular injection of atropine sulfate (Mitsubishi Tanabe Pharma, Osaka, Japan) was administered at 0.04 mg/kg as premedication, followed by an intramuscular injection of a three-mixture solution to induce anesthesia. This solution included 0.4 mg/kg of butorphanol tartrate (Meiji Seika Pharma, Tokyo, Japan), 30 µg/kg of medetomidine hydrochloride (Nippon Zenyaku Kogyo, Fukushima, Japan), and 0.4 mg/kg of midazolam (Astellas Pharma, Tokyo, Japan). After confirming sedation, 6 to 8 mg/kg of propofol (Maruishi Pharmaceutical, Osaka, Japan) was injected intravenously to induce deep anesthesia, after which tracheal intubation was performed. While maintaining anesthesia using 1% to 5% isoflurane inhalation with an artificial ventilator combined with an intravenous drip infusion of propofol, following laparotomy, the peripheral aspect of the greater omentum was isolated by sequential ligation and dissection. Subsequently, multiple thin pieces of the greater omentum were excised. ADMPCs were isolated as per a previous report19
, and the cells were cultured in accordance with the human ADMPC culture method.
, and the cells were cultured in accordance with the human ADMPC culture method.
4
Multimodal Anesthetic Protocol for Canine Surgery
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As premedication, 1.0 mg/kg of maropitant citrate hydrate (Cerenia®; Zoetis, Parsippany, NJ, USA) and 0.04 mg/kg of atropine sulfate (Mitsubishi Tanabe Pharma Co., Osaka, Japan) were administered subcutaneously. In addition, 0.1 mg/kg midazolam (Dormicum®; Astellas Pharma Inc., Tokyo, Japan) and 5.0 μg/kg fentanyl/0.25 mg/kg droperidol (Thalamonal®; Daiichi-Sankyo Propharma Co., Ltd., Tokyo, Japan) were administered intravenously (0.1 ml/kg). For the induction of general anesthesia, propofol (Mylan Seiyaku Ltd., Tokyo, Japan) was intravenously administered, and endotracheal intubation was performed. During the surgery, anesthesia was maintained with isoflurane (IsoFlo®; Zoetis), and artificial ventilation was performed with oxygen.
Continuous intravenous administration of 10–40 mg/kg/h remifentanil hydrochloride (Daiichi-Sankyo Propharma Co., Ltd.) and 25–50 mg/kg/min lidocaine (Aspen Japan, Tokyo, Japan) was also performed for intraoperative analgesia. A 0.3 mg/kg of morphine hydrochloride (Takeda Pharmaceutical Co. Ltd., Osaka, Japan) was administered intramuscularly before and after surgery. After surgery, continuous intravenous administration of 1.25–5.0 mg/kg/h remifentanil hydrochloride (Daiichi-Sankyo Propharma Co., Ltd.) was maintained for postoperative analgesia.
Continuous intravenous administration of 10–40 mg/kg/h remifentanil hydrochloride (Daiichi-Sankyo Propharma Co., Ltd.) and 25–50 mg/kg/min lidocaine (Aspen Japan, Tokyo, Japan) was also performed for intraoperative analgesia. A 0.3 mg/kg of morphine hydrochloride (Takeda Pharmaceutical Co. Ltd., Osaka, Japan) was administered intramuscularly before and after surgery. After surgery, continuous intravenous administration of 1.25–5.0 mg/kg/h remifentanil hydrochloride (Daiichi-Sankyo Propharma Co., Ltd.) was maintained for postoperative analgesia.
5
Dental Infection and Aortic Tissue Analysis
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All surgeries were performed on mice under intraperitoneal anesthesia induced by pentobarbital sodium (1.62 mg/30 g; Kyoritsu Seiyaku Co., Tokyo, Japan) and atropine sulfate (12.5 ug/30 g; Mitsubishi Tanabe Pharma Co., Osaka, Japan). Surgeries were performed in biosafety cabinets of Hiroshima University Animal Facility, and maximum efforts were made to minimize postoperative pain and suffering. To establish dental infection of Pg, the pulp chambers of the maxillary first molars on the left and right sides were opened with a #1/2 round bur. After removing the coronal pulp, a small sterile cotton swab including 1 µl of absorbed bacterial suspension (107 CFU of Pg W83 strain) was inserted into the pulp chamber, which was then sealed with Caviton (GC Co., Tokyo, Japan). Six weeks later, mice were euthanized and samples of maxilla and 5 mm length of aortal tissues from descending aorta at the level equivalent to heart were harvested. Tissues were stored at −80°C for further evaluation or processed as formalin-fixed paraffin embedded (FFPE) samples.
6
Canine Anesthesia and Ventilation Protocol
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All canines were anesthetized with intramuscular injection of ketamine (10 mg/kg; Daiichi Sankyo, Tokyo, Japan), xylazine (0.25 mg/kg; Bayer Yakuhin, Osaka, Japan), and atropine sulfate (0.05 mg/kg; Mitsubishi Tanabe Pharma, Osaka, Japan) and intubated for mechanical ventilation. Anesthesia was maintained via the inhalation of 2–3% sevoflurane (Pfizer, New York, NY, USA) and O2. All donor and recipient canines were ventilated with an inspired O2 fraction of 1.0 at a tidal volume of 20 mL/kg, a respiratory rate of 15 breaths/min, and a positive end-expiratory pressure of 5 cmH2O.
7
Anesthetic Administration and Mitral Valve Repair
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The subjects received subcutaneous administration of atropine sulfate (0.04 mg/kg; Mitsubishi Tanabe Pharma Corp., Osaka, Japan) as a pre-anesthetic
medication. After sufficient oxygenation, anesthesia was induced intravenously with fentanyl (5 µg/kg; Daiichi Sankyo Co., Ltd., Tokyo, Japan)
and propofol (4 mg/kg; Mylan Inc., Canonsburg, PA, USA). After the animals were intubated and placed under inhalational anesthesia with isoflurane (DS Pharma
Animal Health Co., Ltd., Osaka, Japan), we undertook additional procedures to ensure the maintenance of anesthesia via continuous infusion of fentanyl at 5
µg/kg/hr and intravenous administration of vecuronium (0.1 mg/kg; Maruishi Pharmaceutical Co., Ltd., Osaka, Japan). The surgical approach
involved a left lateral thoracotomy via the fifth intercostal space. After cardiac arrest was induced, we performed MVP, specifically mitral chordal replacement
and mitral annuloplasty.
medication. After sufficient oxygenation, anesthesia was induced intravenously with fentanyl (5 µg/kg; Daiichi Sankyo Co., Ltd., Tokyo, Japan)
and propofol (4 mg/kg; Mylan Inc., Canonsburg, PA, USA). After the animals were intubated and placed under inhalational anesthesia with isoflurane (DS Pharma
Animal Health Co., Ltd., Osaka, Japan), we undertook additional procedures to ensure the maintenance of anesthesia via continuous infusion of fentanyl at 5
µg/kg/hr and intravenous administration of vecuronium (0.1 mg/kg; Maruishi Pharmaceutical Co., Ltd., Osaka, Japan). The surgical approach
involved a left lateral thoracotomy via the fifth intercostal space. After cardiac arrest was induced, we performed MVP, specifically mitral chordal replacement
and mitral annuloplasty.
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