Butorphanol Tartrate

Data and samples were collected from wild badgers living in the Woodchester Park study area, a 7 km² region of Cotswold limestone escarpment in Gloucestershire, south-west England (51°43′N, 2°16′W). The resident population of badgers (approximately 300 individuals in 26 social groups) has been the subject of long-term research into badger ecology and TB epidemiology, details of which are given elsewhere [29] . Badgers were captured in the immediate vicinity of their setts in peanut-baited cage traps and transported to a sampling facility to be anaesthetised and examined. All animals were anaesthetised by intramuscular injection of a combination of 8 mg/kg ketamine hydrochloride (Vetalar; Pfizer Ltd, Sandwich, UK), 0.04 mg/kg medetomidine hydrochloride (Domitor; Pfizer Ltd) and 0.8 mg/kg butorphanol tartrate (Torbugesic; Fort Dodge Animal Health, Southampton, UK) [30] . They were then sexed, weighed and measured. On first capture each badger was given a unique identifying tattoo on its ventral abdomen [31] which allowed individuals to be identified thereafter. Samples of faeces, urine, tracheal aspirate, oesophageal aspirate and swabs from bite wounds (where present) were collected for mycobacterial culture and up to 12 ml of jugular blood was taken for serological and gamma interferon testing (see below). After recovery from anaesthesia, badgers were released at the site where they had been captured. Each social group was trapped four times per year. The present study used data derived from 875 capture events that occurred between July 2006 and December 2008, which represented 305 individual badgers (130 male, 175 female) from 26 social groups. Of the badgers caught, individuals were sampled on average three times (range 1 to 10) during the study period. All diagnostic tests gave conclusive results on each of the 875 sampling sessions included in the dataset.

OCT was performed via the previously reported method with slight modifications [25 (link)]. In brief, using a Micron^® IV (Phoenix Research Labs, Pleasanton, CA, USA) with a contact lens specifically designed for mouse OCT, OCT was carried out at 17 time points starting from the postnatal day (P) 15 until P294 for P23H rats and at 6 points from P26 until P247 for SD rats. Although we employed a contact lens designed for rats in previous experiments, we found that the contact lens designed for mice provided a better resolution and clearer images, even for rats, in the OCT [25 (link)]. We therefore employed the contact lens designed for mice in the present study. Two to four rats (four to eight eyes) were evaluated at each time point. In addition, to keep the rats’ cornea sufficiently clear during the OCT examination, we prepared four different age groups of the P23H rats and two age groups of SD rats (three to six rats in each group) and performed OCT measurements alternately using these groups of rats (two to three times per group). Rats were anesthetized with an intraperitoneal injection of a mixture of medetomidine hydrochloride (0.315mg/kg), midazolam (2.0mg/kg), and butorphanol tartrate (2.5mg/kg). To prevent any pain associated with injections, rats were first anesthetized by inhalation of 80% carbon dioxide and 20% oxygen prior to the intraperitoneal injection. The physical conditions, including heartbeat and respiratory pattern, of rats were frequently monitored during experiments by inspection and gentle palpation by the researchers. Pupils were dilated by instillation of a mixture of 0.5% tropicamide and 0.5% phenylephrine hydrochloride eyedrops. Corneal surface was protected using a 1.5% hydroxyethylcellulose solution. The rat ocular fundus was simultaneously monitored using a fundus camera, and the position of the retinal OCT image was set horizontally at 1 disc diameter superior to the optic disc. Fifty images were averaged to eliminate the projection artifacts. The acquired OCT images were quantitatively analyzed using the InSight^® software program (Phoenix Research Labs). Three to eight images from two to five rats each from the both P23H rat and SD rat groups were selected based on the quality of the pictures in terms of the image sharpness at each time point in order to perform as precise a segmentation analysis using InSight^® as possible. The pictures deemed unsuitable for the segmentation analysis because of blurring from rats’ respiratory body movement during OCT experiments were eliminated.

OCT was performed using a Micron^® IV (Phoenix Research Labs, Pleasanton, CA) with a contact lens specifically designed for rat OCT. In the RCS^-/- rats, OCT was conducted at 12 time points between postnatal (PN) day 17 and PN day 111, while in the RCS^+/+ rats, OCT was carried out at eight time points between PN day 18 and PN day 67. At each time point, four to eight rats (eight to 16 eyes) were evaluated. The rats were anesthetized by intraperitoneal injection of a mixture of medetomidine hydrochloride (0.315mg/kg), midazolam (2.0mg/kg), and butorphanol tartrate (2.5mg/kg). To alleviate the pain associated with injection, the rats were pre-anesthetized by inhalation of 80% carbon dioxide and 20% oxygen prior to the intraperitoneal injection. The researchers monitored the physical conditions of the rats including heart beat and respiratory pattern, by inspection and gentle palpation every minute during the experiment. The pupils were dilated using eyedrops that contained a mixture of 0.5% tropicamide and 0.5% phenylephrine hydrochloride. The corneal surface was protected using a 1.5% hydroxyethylcellulose solution. The rat ocular fundus was monitored using the fundus camera of the Micron^® IV, and the position of the retinal OCT image was set horizontally at one disc diameter superior to the optic disc. Fifty images were averaged to eliminate projection artifacts. The acquired OCT images were quantitatively analyzed using the InSight^® software (Phoenix Research Labs). Five images from five rats in each genotype group were selected at each time point on the basis of image sharpness; importantly to avoid selection bias, the pictures were not selected by thickness or reflectivity. We measured the thicknesses of the inner (A, Fig 1), middle (B, Fig 1), and outer (C, Fig 1) layers of the neural retina, as well as that of the combined RPE and choroid (D, Fig 1). The middle layer consists of the combined outer plexiform and outer nuclear layers, and the outer layer consists of the photoreceptor inner segment (IS) and OS layers (Fig 1).

SF samples were obtained by arthrocentesis from meta-carpophalangeal (MCP) joints of 8 horses with OA and 8 horses without joint diseases at the Veterinary Teaching Hospital, University of Helsinki. Most of the horses were of warmblood breeds, but 1 Standardbred and 1 Estonian riding pony were also included. Informed owner consent was obtained for each animal, and ethical approval for SF collection and use was provided by the Viikki Campus Research Ethics Committee of the University of Helsinki (Statement 1/2018). The sampling was conducted immediately after medically-induced euthanasia (0.01–0.02 mg/kg detomidine hydrochloride, 0.01–0.02 mg/kg butorphanol tartrate, 0.05–0.1 mg/kg midazolam, 2.2 mg/kg ketamine hydrochloride, T-61 euthanasia solution) due to lameness or non-OA-related reasons including colic, back pain, leg injury, wound, sinusitis, neurological disease, and guttural pouch mycosis. The decisions to euthanize the animals had been made previously without any relation to the research protocol or sampling. The unprocessed SF samples were immediately frozen in liquid nitrogen and stored at –80°C until analysed. MCP OA was diagnosed post-mortem by experienced equine veterinarians based on the presence of wear lines, erosion of articular cartilage, and osteophytes. Joint surfaces were scored according to OA severity as follows: 0 = normal, 1 = mild OA, 2 = moderate OA, and 3 = severe OA [19 (link)]. SF samples were also harvested from CL MCP joints. Only one of them was classified as normal, and the rest had mild-to-moderate OA. In the post-mortem examination of control joints, the MCP joint surfaces either had no macroscopic abnormalities or revealed mild periarticular changes. Regarding medication, 3 control and 2 CL/OA horses had been treated with nonsteroidal anti-inflammatory drugs, 1 control and 1 CL/OA horse had received antibiotics, and 1 CL/OA horse had been treated with antifungals.

Bovine Collagen type I, 8 wt.% aqueous solution (Coll, Collado s.r.o., Brno, Czech Republic), chitosan from shrimp shells, 70% DDA, low viscosity (Chit, Sigma-Aldrich, Darmstadt, Germany), calcium salt of oxidized cellulose–degree of oxidation 16–24% and M_n = 350 kg/mol (CaOC, Synthesia, Pardubice, Czech Republic), acetic acid (99%, Penta s.r.o, Chrudim, Czech Republic), poly(ε-caprolactone) (PCL, 80 kg/mol), gelatin (Gel, Type B, Bioreagent, powder from bovine skin), N-(3-Dimethylaminopropyl)-N´-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), 98% dopamine hydrochloride, tris (hydroxymethyl) aminomethane hydrochloride, ethanol p.a. 99.8%, sodium phosphate dibasic for molecular biology (≥ 98,5%), sodium chloride, calcium chloride, sodium phosphate dibasic dodecahydrate (Na₂HPO₄ ·12H₂O), potassium dihydrogen phosphate (KH₂PO₄), potassium chloride (KCl), collagenase from Clostridium histolyticum, lysozyme human, the murine fibroblast cell lines 3T3-A31, Dulbecco's modified eagle medium DMEM (D6429), fetal bovine serum FBS (F7524), 2′,7′-bis (2-carboxyethyl)-5(6)-carboxyfluorescein acetoxymethyl ester (BCECF-AM), propidium iodide (P4864), (all from Sigma Aldrich, Darmstadt, Germany), penicillin/streptomycin (15140–122) and DiOC6(3) (D273), (Life Technologies, Eugene, OR, USA), octenidine solution (Octenisept®, Schülke, Germany), Butomidor^® inj. (butorphanol tartrate, Vétoquinol, Czech republic), Domitor^®, Medetomidine, Orion corporation, Finland) Propofol^® (Propofolum 1%, Fresenius Kabi Deutschland, Bad Homburg, Germany), Metacam^® (meloxicam, Boehringer Ingelheim Vetmedica, Ingelheim/Rhein, Germany), Enroxil^® (Enrofloxacin, Krka, Novo mesto, Slovenia) Betadine^®, (2.5% solution of povidone iodine, EGIS Pharmaceuticals PLC, Budapest, Hungary) were used as received without further purification.

A murine LIM model was prepared as previously reported^{40 (link)}. We created a mouse eyeglass frame that conformed to the contour of the mouse's head and printed it out using a three-dimensional printer. A negative 30 D lens made of PMMA was created for myopia induction. Myopic induction using the − 30 D lens showed greater myopic shift compared to the form-deprivation myopic model^{40 (link)}. With some differences from the LIM model used previously, we used binocular myopic induction instead of monocular induction. The left and right eyes of the glasses were adjusted by the shape of the mouse skull frame and fixed on the stick with a screw, and then glued the Stick to the mouse skull with a self-cure dental adhesive system. This was done under general anesthesia with the combination of midazolam (Sandoz K.K., Minato, Japan), medetomidine (Domitor®, Orion Corporation, Turku, Finland), and butorphanol tartrate (Meiji Seika Pharma Co., Ltd., Tokyo, Japan) (MMB). The dosage for each mouse was 0.01 ml/g.
During the myopia induction phase, mice were given either normal (MF, Oriental Yeast Co., Ltd, Tokyo, Japan) or mixed chow containing the candidate chemical 0.0667 percent GBEs (INDENA JAPAN CO., Tokyo, Japan #9,033,008). 0.0667% GBEs contain 24% of the flavonol glycosides of quercetin, kaempferol, and isorhamnetin and 6% terpene trilactones. The corresponding concentration of GBEs mixed chow was 200 mg/kg/day, which is consistent with the concentration of GBEs that causes the significantly high activity of EGR-1 in vitro experiments. The addition of GBEs and the production of 0.0667% GBEs mixed chow are all produced by chow manufacturing company (Oriental Yeast Co., LTD., Tokyo, Japan).