Clinical and basic scientists at academic medical and biomedical research institutions often form ideas that could have both monetary and human health benefits if developed and applied to improvement of human wellbeing. However, such ideas lose much of their potential value in both regards if they are disclosed in traditional knowledge-sharing forums such as abstracts, posters, and oral presentations at research meetings. Learning the basics about intellectual property protection and obtaining professional guidance in the management of intellectual property from a knowledgeable technology management professional or intellectual property attorney can avoid such losses yet pose a minimal burden of confidentiality on the investigator. Knowing how to successfully navigate the early stages of intellectual property protection can greatly increase the likelihood that discoveries and knowledge will become available for the public good without diminishing the important mandate of disseminating knowledge through traditional knowledge-sharing forums.

A novel therapeutic compound was found to induce bladder tumors in male rats. Given the location of the tumors and the increased amounts of calcium- and magnesium-containing solids found in the urine of treated animals, we hypothesized that tumorigenesis was secondary to urine crystal formation rather than a direct effect of the drug on urothelium. To investigate the basis for the response, a method of acidifying rodent urine was needed. This study tested the efficacy of 1% dietary NH₄Cl in reducing the urinary pH of male mice. After 1 wk, urinary pH (mean ± SD) at 1 h after light onset was 7.51 ± 0.32 among controls compared with 6.21 ± 0.31 for the NH₄Cl-fed group. After 2 wk of supplementation, urinary pH was 7.78 ± 0.41 for controls and 6.20 ± 0.30 for the NH₄Cl-fed group. To investigate whether the time of collection altered urinary pH, samples also were collected 8 h after the start of the light cycle on the day of the 2-wk collection. Urinary pH was 7.12 ± 0.28 for the control group and 5.80 ± 0.23 for the NH₄Cl-fed mice. The pH differences between control and NH₄Cl-fed groups and the differences in pH within groups at 1 and 8 h were statistically significant. Dietary NH₄Cl is an effective urinary acidifier for mice. When evaluating the pH of mouse urine, care should be taken to compare samples collected at the same time after the start of the light cycle.

The purposes of this study were to determine 1) whether magnetic resonance imaging (MRI)-based T2 mapping and measurements of limb volume can differentiate injured and uninjured tissue after blunt trauma to rat hindlimbs and 2) whether administration of buprenorphine influences these assessments. Male Wistar rats (age, 3 to 4 mo) underwent blunt contusion injury to the posterior aspect of the hindlimb; MRI was conducted at 6, 12, 24, 48, 72, and 96 h after injury. The imaging results showed that administration of buprenorphine had no effect on the T2 value {area under the receiver operating characteristic [ROC] curve: with drug, 0.869 [95% confidence interval (CI), 0.78 to 0.96]; without drug, 0.809 [95% CI, 0.72 to 0.90]} but did influence limb volume [area under the ROC curve; without drug, 0.954 (95% CI, 0.92 to 0.99); with drug, 0.713 (95% CI, 0.61 to 0.82)]. When using MRI to determine the extent of injury or to track injury over time, calculated limb volumes may lose sensitivity to detect injury, due to the intrinsic increase in volume from morphine-derived drugs. During administration of morphine derivatives, T2 maps may provide more accurate assessments of muscle tissue injury both initially after injury and over time.

Determination of the proportion of neutrophils in the peripheral blood is important for diagnostic purposes in medicine and for evaluating new drugs in the pharmaceutical industry. To measure the neutrophil concentration in rat blood, a fast and accurate flow cytometric method was developed. Rat neutrophils were quantified by using primary antibodies that recognize the RP1 antigen and secondary antibodies conjugated with fluorescein isothiocyanate. The flow cytometric method was calibrated by comparing cytometric results with data from a manual differential count. The results obtained by these 2 methods correlated with a Pearson correlation coefficient of 0.91 and were in agreement according to subsequent statistical analysis. To confirm the usefulness of the method in preclinical applications, the production of neutrophils in rats was stimulated by pegfilgrastim. Blood samples were taken at predetermined time points, and the pharmacodynamic profile was determined. These results confirmed that the flow cytometric method for neutrophil quantification is accurate and much faster than the manual microscopic method. Moreover, the flow cytometric method is easy to use, suggesting that it could become the method of choice for preclinical applications.

Cage change and gavage are routine procedures in animal facilities, yet little is known about whether housing modifications change responses to these procedures. Telemetric activity and cardiovascular parameters were assessed in this experiment. BN and F344 male rats were housed in open or individually ventilated cages, each containing 3 rats, 1 of which had a transponder. A crossover design was used, in which 2 groups were given dividers made of 2 intersecting boards (1 form contained holes loaded with food pellets; the other did not) and 1 group was given a rectangular tunnel. On day 8 of each 2-wk period, the cages were changed; on day 11, rats were gavaged. The parameters were evaluated for the first hour and for the following 17 h. Baseline values for each rat were subtracted from the corresponding response values. The presence of objects did not affect the responses of F344 rats to cage changing or gavage. In BN rats with IVCs, the presence of the plain divider modified the response to both procedures. Responses to procedures appeared to be dependent on both the strain and the cage object, thus complicating the establishment of valid general recommendations.

Although nasotracheal intubation in the rabbit has been briefly described, scientific assessment of the procedure has not been reported. In this report we describe nasotracheal intubation performed in 38 male New Zealand White rabbits (3.0 to 5.5 kg) used for a vascular patch study. The rabbits were placed under general anesthesia twice, with 2 mo between the initial and final intubations. Rabbits were intubated by the oral or nasotracheal route and compared. Previous literature dismissed nasotracheal intubation, citing the possibility of introducing pathogens into the lungs and the necessity of high oxygen flow rates (presumably greater than 3 L/min). However, no clinical signs of respiratory disease were noted among the study animals, nor were high oxygen flow rates necessary. Several key points collectively facilitated a successful procedure. Total relaxation was essential, modification of the classic blind approach eased placement, a correct approach was necessary, and our development of a unique method of securing the tube improved tube management. The findings suggest that nasotracheal intubation can be used as an easy, less traumatic method of rabbit intubation when compared with orotracheal intubation.

The experimental use of amphibian models in biomedical research increases yearly, but there is a paucity of reports concerning analgesic use in many of these species. In this study, buprenorphine given by intracoelomic injection and butorphanol added to the tank water were compared for analgesic effect in the eastern red-spotted newt after bilateral forelimb amputations. Newts undergoing anesthesia but not surgery and newts having surgery but not given analgesia postoperatively were used as control groups. Animals were tested for food consumption, spontaneous movement, response to tapping on the tank, response to being touched, and body posture. Both buprenorphine by intracoelomic injection and butorphanol in tank water significantly promoted resumption of normal behavior after bilateral surgical amputation of the forelimbs. The difference between analgesic treatment and no analgesic treatment was maintained until 72 h after surgery.

The short-term effects on rates and durations of self-injurious behavior and self-directed stereotypies associated with various doses of fluoxetine (FLX) and venlafaxine (VEN) were examined in rhesus macaques. Adult male macaques (Macaca mulatta; n = 17; age, 7 to 15 y) with at least 1 episode of severe SIB within the past 5 y were randomized to treatment with FLX (n = 6), VEN (n = 6), or placebo (PLC, n = 5), administered by voluntary consumption of medication provided in fruit-flavored tablets. After 4-wk baseline and 4-wk placebo lead-in phases, doses were increased monthly for 4 mo (FLX: 0.5, 2.0, 4.0, and 8.0 mg/kg; VEN: 2.0, 4.0, 8.0, and 16.0 mg/kg). Animals in the PLC condition received similar nonmedicated fruit-flavored tablets. Focal behavioral observations, plasma drug levels, and neurochemical data were obtained. Results indicated that rates and percentage time spent self-biting declined at all doses of FLX, with the greatest effect seen at 2.0 mg/kg. For VEN, percentage time spent self-biting was significantly lower only at the 4.0 mg/kg dose. Treatment-induced reductions in platelet serotonin and cerebrospinal fluid 5-hydroxyindoleacetic acid (CSF 5HIAA) concentrations were substantially greater in the FLX-treated condition than in the VEN-treated condition. Plasma FLX and norfluoxetine levels increased with FLX dose; plasma levels of VEN were low and not dose-related. Fluoxetine at a dose of 2.0 mg/kg daily was most efficacious in reducing SIB, and the observed reductions in platelet serotonin and CSF 5HIAA levels indicated substantial bioeffect at this dose. Treatment with VEN was marked by noncompliance, low bioeffect, and low efficacy.

Self-injurious behavior (SIB) among captive primates is a recurring problem for those who manage such facilities. Its prevalence highlights the need for research evaluating the effectiveness of potential treatment approaches. In the present study, 4 wk of dietary supplementation with L-tryptophan (100 mg daily) was evaluated for the treatment of self-inflicted wounds in 22 small-eared bushbabies, a prosimian primate, with a history of SIB. The treatment significantly reduced stereotypy and was associated with a reduction in wound area and severity. In terms of physiologic measures, preexisting high levels of cortisol were reduced in bushbabies with SIB, whereas serotonin concentrations were increased after 4 wk of treatment. Results indicate that L-tryptophan as a dietary supplement may be a viable adjunct to standard husbandry procedures for animals exhibiting maladaptive behaviors such as stereotypy and SIB.

Positive reinforcement training (PRT) has successfully been used to train diverse species to execute behaviors helpful in the everyday care and wellbeing of the animals. Because little information is available about training sooty mangabeys (Cercocebus atys atys), we analyzed PRT with a group of 30 adult males as they were trained to shift from 1 side of their enclosure to the other. Over a 4-mo period we conducted 57 training sessions totaling 26.5 h of training and recorded compliance information. During training, compliance increased from 76% of the animals during the first 5 training sessions to 86% of the animals shifting during the last 5 sessions. This result indicated progress but fell short of our goal of 90% compliance. After 25 training sessions, problem-solving techniques were applied to help the consistently noncompliant animals become more proficient. The techniques included reducing social stress by shifting animals so that noncompliant monkeys could shift into an unoccupied space, using more highly preferred foods, and 'jackpot'-sized reinforcement. To determine whether social rank affected training success, animals were categorized into high, medium, and low dominance groups, based on 7 h of behavioral observations. A Kruskal–Wallis test result indicated a significant difference in compliance according to the category of dominance. Although training a group this large proved challenging, the mangabeys cooperated more than 90% of the time during follow-up sessions. The training program improved efficiency in caring for the mangabeys.

Assessment of fecal glucocorticoid metabolites has become a widely used method for monitoring stress responses. Because most small rodents are social animals whose physiologic parameters are affected by social stimuli, individual housing may compromise these data. Nevertheless, housing rodents in families or social groups may be an important limitation to the experimental design. The challenge is to collect samples from individual rodents while avoiding stress-associated effects from the sampling method itself. Here we present an apparatus and protocol allowing routine repeated collection of an individual rodent's fresh fecal samples without noticeable disturbance of any of the study animals; continuous maintenance of studied animals in a familiar environment; group housing; and uninterrupted visual and olfactory communication among group members during sampling. The apparatus consists of 1 central and 4 lateral compartments. The experimental animal was allowed to enter a lateral compartment voluntarily, where it remained for the short (4 h) period necessary for sample collection before rejoining the rest of the group. Evaluations involved Egyptian spiny mice, a social rodent increasingly studied in laboratories. The results confirmed the repeatability of the assessment of baseline levels of glucocorticoid metabolites. Moreover, keeping the animals in our experimental apparatus did not induce any increase in the levels of glucocorticoid metabolites, even when isolation in the compartment was relatively prolonged. We interpret these results as confirmation that our sampling procedure allows repeated individual sampling within a nearly undisturbed social unit.

Serum is often frozen and banked for analysis at a later date. This study assessed the stability of 17 analytes in rat serum during refrigeration at 4 °C and extended storage at −20 °C (frost-free and nonfrost-free freezers) and −70 °C. Samples were analyzed by using an automated dry-slide chemistry analyzer at time 0 and then stored as aliquots for analysis at time points including day 7, 30, 90, and 360. After 7 d of refrigeration, only creatine kinase activity had varied by more than 10% of the starting value. Freezing at −70 °C was clearly superior to −20 °C where changes were observed in CO₂ as early as day 30 and alanine aminotransferase as early as day 90. Samples stored in frost-free and nonfrost-free −20 °C freezers did not differ significantly through day 90. Factors such as storage time and temperature should be considered when designing any retrospective study.