Safety and immunogenicity studies were performed in New Zealand White rabbits (Envigo, Madison, WI, USA). Administration of the test articles, in-life monitoring, and necropsy (including organ harvest and serum collection) were conducted by Frontage Laboratories, Inc. (Concord, OH, USA). The age of animals at the time of dosing was 4–5 months. All animals were experimentally naïve (10 males and 10 females). The placebo and VaxiStrep test articles were provided by VaxForm as ready-to-inject formulations. All administrations were given via intramuscular injection (IM) in the right hindlimb of the rabbits. Placebo or VaxiStrep were administered to 10 rabbits each (5 male and 5 female), on days 1 and 29 at 0 or 25 µg/animal/dose, respectively, at a dose volume of 0.5 mL/animal to mimic the human dose. Blood samples for immunogenicity testing were collected from all animals during the pre-dose period (day—1), and on days 15, 22, 29, and 33. The animals were terminated on day 33. The heart, kidneys, and brain were collected from all terminated animals as whole organs and preserved in 10% neutral-buffered formalin for immunohistochemistry.
New zealand white rabbits
Manufactured by Inotiv
Sourced in United States
New Zealand White rabbits are a breed of domestic rabbit commonly used in laboratory research. They are a medium-sized breed with a white coat and pink eyes. New Zealand White rabbits are known for their docile temperament and are often used as a standard model organism in various scientific studies and experiments.
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6 protocols using new zealand white rabbits
1
Safety and Immunogenicity of VaxiStrep in Rabbits
2
Nonischemic Heart Failure Rabbit Model
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Nonischemic HF was induced in male New Zealand White rabbits (Envigo, Indianapolis, IN, USA, and Charles River, Wilmington, MA, USA) by combined volume and pressure overload [3 (link),18 (link)]. Volume overload was induced by surgical creation of aortic valve insufficiency, followed 2–4 weeks later by induction of pressure overload by abdominal aortic banding. Surgeries were conducted while the rabbits were under isoflurane inhalation anesthesia after premedication with a combination of ketamine, xylazine and acepromazine. HF developed gradually (over 5–8 months, monitored by serial echocardiography) and was characterized by depressed systolic ventricular function. Atrial cells were isolated from hearts that showed marked left-ventricular dilation (54 ± 5% increase in left ventricular diastolic internal dimension) and systolic dysfunction (decrease of left-ventricular fractional shortening by 37 ± 4%). This HF rabbit model has been extensively studied in ventricular and, to some extent atrial myocytes and has been previously characterized at structural, molecular, Ca2+ handling, and electrophysiological levels [3 (link),18 (link),19 (link),20 (link)]. In this study 19 HF rabbits and 87 normal rabbits were used.
3
Rabbit Hypertrophic Scar Model
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Animal experiments were approved by the Institutional Animal Care and Use Committee at Northwestern University. All experiments were performed in female, 3‐5 kg New Zealand White rabbits (Envigo, Indianapolis, IN). The hypertrophic scar (HTS) model was performed as described previously.6 , 7 Briefly, rabbits were anesthetized with ketamine/xylazine, and the ventral side of each ear was shaved and depilated. To each ear, six full‐thickness circular excisional wounds of diameter 7 mm were created down to the perichondrium, and excised tissue removed using forceps (Figure 1A ). Wounds were covered with Tegaderm (3M Healthcare, St. Paul, MN) and rabbits were kept until POD14 to allow wound re‐epithelialization. Application of creams (vehicle or drug formulations) began on POD14 and was performed 5x per week for 2 weeks. Wounds were harvested at POD28 for analysis unless otherwise specified. Trans‐epidermal water loss (TEWL) was measured using a Dermalab Combo and TEWL probe (cyberDERM, inc. Media, PA).
4
Pharmacokinetics of BDF in Rats and Rabbits
5
Polyclonal Antibodies Production against OMVs
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To produce specific polyclonal antibodies against NT-OMV and HT-OMV sediment, New Zealand White rabbits (Envigo, Indianapolis, IN, USA) were immunized with 1 mg of the corresponding antigen in 0.2 mL of PBS, intramuscularly. Blood samples were collected before immunization and weekly until four weeks post-immunization. The presence of specific antibodies against bacterial vesicles in serum were determined by immunoblotting at week 0, 1, 2, 3, and 4 post-immunization. Immunoblotting was performed using nitrocellulose membranes (Whatman Protran®; Merk kGaA, Darmstadt, Germany, pore size 0.45 µm) and a semidry blotting system at 0.8 mA/cm2 for 30 min (Trans-Blot® SD Transfer Cell, BIO-RAD, Hercules, CA, USA). After that, protein-binding sites were blocked with PBS with 5% skimmed milk, overnight, at room temperature. Then, the membranes were washed 3 times with PBS-Tween and then incubated (4 h at 4 °C) with sera from hyperimmunized rabbits collected at week 0, 1, 2, 3, and 4 post-immunization. The membranes were washed 3 times with PBS-Tween and then peroxidase-conjugated secondary antibody (anti-IgGFc,) was added for 60 min at room temperature. Finally, membranes were washed with PBS-Tween and the antibody-antigen complexes were visualized after addition of a substrate/chromogen solution (H2O2/chloro α-naphthol).
6
Standardized Animal Handling and Welfare Protocols
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Male Lister hooded rats, C57BL/6J mice, and New Zealand white rabbits were purchased from Envigo. Brown Norway male rats were sourced from Charles River. Animals were acclimated for at least 7 days and housed in a pathogen-free setting with 12-hour light–dark cycles and controlled temperature (22 ± 1 °C) and humidity (55 ± 5%) ranges and access to food and water ad libitum. Animals were cared for by trained personnel, and procedures were conducted by trained, licensed scientists. All procedures were conducted in accordance with the United Kingdom Animals (Scientific Procedures) Act 1986 and approved by the Department of Health, Social Services, and Public Safety of Northern Ireland. For laser CNV experiments performed by Iris Pharma (La Gaude, France), all procedures and protocols were reviewed by an internal ethics committee (identifier: DAP53), and animals were treated according to Directive 2010/63/EU of the European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes. All experiments were conducted in compliance with the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmology and Vision Research.
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