Ketanest s

Male C57BL/6 mice aged 10–12 weeks were obtained from Janvier Laboratories (Le Genest Saint Isle, France). Mice were housed in a 12-h light/dark cycle at 22 °C. Food and water were provided ad libidum. Animals were allowed to acclimatize for 7 days before any experimental procedure. Polymicrobial sepsis was induced using the cecal ligation and puncture (CLP) model [52 (link)]. In total, 20 mice were anesthetized with 100 mg/kg ketamine (Ketanest®S, Pfizer Pharma, Berlin, Germany) and 20 mg/kg xylazine (Xylavet, CP-Pharma, Burgdorf, Germany) intraperitoneally. After a midline laparotomy and mobilization of the cecum, 5 mm was ligated and punctured once with a 23-G needle (BD Microlance™ 3, BD Medical, Heidelberg, Germany). The ligated cecum was pressed gently to extrude fecal contents. Afterwards, the cecum was relocated, the mice were supplemented with 400 μl 0.9% NaCl (B. Braun, Melsungen, Germany), given directly into the abdominal region, and the abdomen was closed with a double suture. For control, 10 animals underwent a laparotomy surgery only applying the same anesthesia regime. After surgery, pain relieve in both groups was achieved by treatment with 0.05 mg/kg bodyweight buprenorphine (Temgesic, RB Pharmaceuticals, Slough, UK) every 8 h for 2 days. For the isolation and analysis of sperm, a separate cohort of animals, both CLP as well as sham, was operated.

For the investigation of post-septic immunological consequences a cecal ligation and puncture (CLP) mouse model was used as described before (21 (link)). Briefly, mice were anesthetized by an intraperitoneal injection of 100 mg/kg ketamine (Ketanest®S, Pfizer Pharma, Berlin, Germany) and 20 mg/kg xylazine (Xylavet, CP-Pharma, Burgdorf, Germany). After median laparotomy, cecum was mobilized, ligated (5 mm).and punctured once with a 23 G needle (BD Microlance™ 3, BD Medical, Heidelberg, Germany). Fecal content was gently extruded and the cecum afterwards relocated. Mice were supplemented with 400 μL 0.9% sodium chloride (B. Braun, Melsungen, Germany), given directly in the abdominal cavity and abdomen was closed thereafter with a double suture. Control animals received a sham surgery without cecal ligation and puncture. For pain relieve, mice were treated with 0.05 mg/kg bodyweight buprenorphine (Temgesic, RB Pharmaceuticals, Slough, UK) every 8 h for 2 days after surgery. In total, 15 animals received a CLP and 9 animals a sham surgery. All animals were subsequently housed till euthanasia 12 weeks after intervention (Figure 1A).

All rats were obtained from our colonies at the Forschungseinrichtung für experimentelle Medizin (FEM), Charité—Universitätsmedizin, Berlin. SHRSP rats and WKY rats from these colonies were previously described [18 (link),19 (link)] and breeder pairs of F344 rats were obtained from Charles River (Charles River Laboratories International, Inc.) to establish a new colony at our facility. The total number of male animals studied were for SHRSP E20 n = 7, week 14 n = 8, for F344 E20 n = 8, week 14 n = 8, and for WKY E20 n = 8, week 14 n = 17. Rats were grouped under a 12:12h light/dark cycle using an automated light switching device and climate-controlled conditions at 22°C. Adult rats at week 14 and maternal animals were fed a normal-salt diet (0.2% NaCl). All animal experiments were approved by the government committee in accordance with national animal protection guidelines (Landesamt für Gesundheit und Soziales (LAGeSo) Berlin, Germany).
At week 14 of age animals were weighed and sacrificed under a ketamine (Ketanest S, Pfizer, Karlsruhe, Germany)/xylazine (Rompun, Bayer, Leverkusen, Germany) anaesthesia (87 mg/kg and 13 mg/kg body weight, respectively).
Fetuses on stage E20 were dissected from the uterus and immediately killed by decapitation. Maternal animals were anesthetized by inhalation of isoflurane (Abbott, Wiesbaden, Germany) and sacrificed by removing the heart.

Male New Zealand white rabbits (Harlan, Netherlands B.V.) were housed individually in polycarbonate cages (Scanbur Number 8 plastic cages with stainless steel door) with aspen bedding (Aspen Bricks M, Tapvei Ky, Kaavi, Finland). The acclimation period before experiments was at least 21 days. Rabbits weighing 2.0–2.3 kg (age 10–12 weeks) were used for haemodynamically assessing ORM‐11372 (no randomization, baseline values as own control). The diet provided to rabbits was SDS Stanrab (P) SQC pelleted (Special Diet Services Ltd, Witham, England). Water was provided ad libitum in polycarbonate bottles (750 ml).
Rabbits (n = 5) were sedated with i.v. (marginal vein) diazepam (2 mg·kg⁻¹, Diapam®, Orion Pharma). Anaesthesia was induced with i.v. S‐ketamine (10–20 mg·kg⁻¹ Ketanest‐S®, Pfizer) and maintained with S‐ketamine i.v. infusion (15–80 mg·kg⁻¹·h⁻¹). Animals were placed on a heating table (+38°C), and tracheas were cannulated. Rabbits were ventilated via a rodent ventilator (Ugo Basile 7025, Hugo Sachs Elektronik, Germany; respiratory volume 10 ml·kg⁻¹, 30 strokes·min⁻¹ for rabbits). ORM‐11372 was infused into the jugular vein at an infusion rate of 10 ml·kg⁻¹·h⁻¹ (Terumo TE‐311, Belgium), at infusion doses of 17, 167, and 833 μg·kg⁻¹·min⁻¹.

This randomised, open-label, controlled, parallel group, Phase IV clinical drug trial (ClinicalTrials.gov identifier NCT02624401) was conducted at Turku PET Centre, University of Turku, Turku, Finland as a part of ‘The Neural Mechanisms of Anesthesia and Human Consciousness’ project (from January 2016 to March 2017), as predefined in the trial protocol. This study was approved by the Ethics Committee of the Hospital District of Southwest Finland and the Finnish Medicines Agency Fimea (EudraCT 2015-004982-10). This article adheres to the applicable Consolidated Standards of Reporting Trials (CONSORT) guidelines. A detailed description of the study methods and the CONSORT flow diagram have been published earlier.4 (link), 7 (link)
A total of 160 healthy, ASA physical status Class 1 male subjects were randomly allocated to receive one of the following study treatments: dexmedetomidine (Dexdor 100 μg ml⁻¹; Orion Pharma, Espoo, Finland; n=40), propofol (Propolipid 10 mg ml⁻¹; Fresenius Kabi, Uppsala, Sweden; n=40), sevoflurane (Sevoflurane 100%; AbbVie, Espoo, Finland; n=40), S-ketamine (Ketanest-S 25 mg ml⁻¹; Pfizer, Helsinki, Finland; n=20), or saline placebo (n=20). The inclusion criteria have been described earlier.⁷ (link) In accordance with the Declaration of Helsinki, a written informed consent was obtained from all study subjects.