Effects of Glucagon-Like Peptide-1 Receptor Agonist Exendin-4 on the Reinstatement of Cocaine-Mediated Conditioned Place Preference in Mice

A high percentage of relapse to compulsive cocaine-taking and cocaine-seeking behaviors following abstinence constitutes a major obstacle to the clinical treatment of cocaine addiction. Thus, there is a substantial need to develop effective pharmacotherapies for the prevention of cocaine relapse. The reinstatement paradigm is known as the most commonly used animal model to study relapse in abstinent human addicts. The primary aim of this study is to investigate the potential effects of systemic administration of glucagon-like peptide-1 receptor agonist (GLP-1RA) exendin-4 (Ex4) on the cocaine- and stress-triggered reinstatement of cocaine-induced conditioned place preference (CPP) in male C57BL/6J mice. The biased CPP paradigm was induced by alternating administration of saline and cocaine (20 mg/kg), followed by extinction training and then reinstatement by either a cocaine prime (10 mg/kg) or exposure to swimming on the reinstatement test day. To examine the effects of Ex4 on the reinstatement, Ex4 was systemically administered 1 h after the daily extinction session. Additionally, we also explored the associated molecular basis of the behavioral effects of Ex4. The expression of nuclear factor κβ (NF-κβ) in the nucleus accumbens (NAc) was detected using Western blotting. As a result, all animals that were treated with cocaine during the conditioning period successfully acquired CPP, and their CPP response was extinguished after 8 extinction sessions. Furthermore, the animals that were exposed to cocaine or swimming on the reinstatement day showed a significant reinstatement of CPP. Interestingly, systemic pretreatment with Ex4 was sufficient to attenuate cocaine- and stress-primed reinstatement of cocaine-induced CPP. Additionally, the expression of NF-κβ, which was upregulated by cocaine, was normalized by Ex4 in the cocaine-experienced mice. Altogether, our study reveals the novel effect of Ex4 on the reinstatement of cocaine-induced CPP and suggests that GLP-1R agonists appear to be highly promising drugs in the treatment of cocaine use disorder.

Carcinogenicity Evaluation of Baclofen in TgrasH2 Mice

Baclofen is a γ-aminobutyric acid-B receptor agonist used for control of spastic muscle activity and as a treatment for alcohol abuse. The review of the nonclinical database suggested a data gap for potential carcinogenicity following long-term use. Regulatory requirements for pharmaceutical safety testing of cancer-causing potential have historically included 2-year rodent studies in rats and mice. The availability of transgenic models with greater specificity and sensitivity to carcinogens provides safety testing alternatives that align with the 3Rs. The carcinogenicity of baclofen was evaluated in CB6F1-TgrasH2 transgenic mice following daily oral administration at 45, 90, and 180 mg/kg/d for 26 weeks, preceded by a 2-week drug-conditioning period. There were no treatment-related palpable masses or neoplastic findings, and survival rates were not affected by the baclofen treatment. In conclusion, baclofen was considered as noncarcinogenic in CB6F1-TgrasH2 mice, which is consistent with results previously obtained in a 2-year rat study.

Can Lambs in Pain Identify Medicated Feed?

Lambs in Australia undergo painful husbandry procedures as part of common husbandry. The magnitude and duration of pain are difficult to assess in lambs. Most currently used methods rely on behavioral expressions and physiological markers that may fail to detect the state of pain an animal experience. This study examined motivation of 12-week-old lambs experiencing chronic pain to self-medicate by consumption of feed containing an analgesic agent as an indicator of pain in lambs. In this study, 36 male Merino lambs were individually penned and acclimated to pelleted feed and two artificial odors: strawberry and banana. Once acclimated to odored feed, lambs were tested for their individual preference for the odors. Lambs were then assigned to one of two groups: Sham—sham handled day 0 and 7 or Ring—Ring castrated day 0 and tail docked day 7. To enable self-medication testing, lambs underwent a conditioning period (day 0–3) followed by the self-medication period (day 7–12). On day 0 lambs were castrated or sham handled, and then offered only medicated feed that contained an odor cue (either strawberry or banana). On day 7, lambs underwent tail-docking or sham handling and were offered both the conditioned medicated feed and non-medicated feed. Amount of each feed consumed was recorded 1 and 12 h after offer each day. Blood samples were taken for cortisol and white blood cell analysis and behavioral observations were recorded for 12 h following treatment. There was no difference in preference for medicated feed between Ring and Sham lambs during the self-medication phase (P = 0.18). Lambs in both groups displayed a significant preference for strawberry cued medicated feed during the self-medicated period when compared to the other testing periods (P = 0.05). Ring lambs displayed more active pain behaviors (mean = 15.1) than Sham (mean = 0.4, P < 0.05). Following castration, Ring lambs had a higher neutrophil/lymphocyte ratio at 6, 24, 48, and 72 h. This study was not able to demonstrate that lambs can self-medicate for a state of pain.

Differential Aspects of Natural and Morphine Reward-Related Behaviors in Conditioned Place Preference Paradigm

Natural rewards are essential for animal survival. On the other hand, drug-seeking behaviours can be maladaptive and endanger survival. The present study was conducted to enhance our understanding of how animals respond to food and morphine as natural and drug rewards, respectively, in a conditioned place preference (CPP) paradigm. We designed a protocol to induce food-CPP and compare it as a natural reward with morphine-CPP in rats. The protocol for reward induction in all groups (foods and morphine) consisted of three phases: pre-test, conditioning, and post-test. In morphine groups, we injected morphine as a reward (5 mg/kg, s.c.), and for inducing natural-reward, we used two different protocols, by one of the rats were deprived of food for 24h. In the other method, rats were restricted for food for 14 days. During the conditioning period, animals received daily chow, biscuit, or popcorn as a reward inducer. Results revealed that CPP did not induce in food-deprived rats. A combination of food restriction (as a facilitator) and a biscuit or popcorn induced reward using CPP. In contrast, food deprivation did not facilitate food-CPP in response to regular food. Interestingly the CPP score of the group which received biscuit during a 7-day conditioning period was more than that of the morphine group. In conclusion, food restriction could be a better protocol than food deprivation to facilitate food reward.

Water-Soluble Carbohydrate Recovery in Pastures of Perennial Ryegrass (Lolium perenne L.) and Pasture Brome (Bromus valdivianus Phil.) Under Two Defoliation Frequencies Determined by Thermal Time

The objectives of the experiment were to (i) examine the dynamics of WSC use and the recovery of leaf sheaths and blades of Bromus valdivianus Phil. and Lolium perenne L. subjected to two defoliation frequencies (DFs) determined by thermal time (TT); (ii) evaluate how DF influenced regrowth and accumulated herbage mass (AHM) during fall. Defoliation was carried out at frequencies of 135 and 270 accumulated growing degree days (AGDDs) for both species. Twelve plots were arranged in a three-block design. All plots had a conditioning period to establish the assigned DF prior to sampling. From the start of the experiment, “cores” were collected from each plot every three days until the DF was reached. Every core was separated into leaf and sheath material before measuring the WSC concentration. Lolium perenne had concentrated more WSCs than B. valdivianus. Both species adapted their WSC recovery according to the DF. The recovery of WSC was faster under a DF of 135 AGDDs than that of 270 AGDDs. Leaf sheaths contained more WSCs than leaf blades and were identified as WSC storage organs. This period can be used as the optimal defoliation interval in B. valdivianus and L. perenne grazing systems.

Fear Conditioning Potentiates the Hippocampal CA1 Commissural Pathway In vivo and Increases Awake Phase Sleep

The hippocampus is essential for spatial learning and memory. To assess learning we used contextual fear conditioning (cFC), where animals learn to associate a place with aversive events such as foot-shocks. Candidate memory mechanisms for cFC are long-term potentiation and long-term depression, but there is little direct evidence of those mechanisms operating in the hippocampus in vivo following cFC. Also, little is known about the behavioral state changes induced by cFC. To address these shortcomings, we used motion tracking and recorded local field potentials in freely behaving mice by placing a stimulating electrode in the left dorsal CA1 region and positioning recording electrodes in the right dorsal CA1 region. Synaptic strength in the commissural pathway was monitored by measuring field excitatory postsynaptic potentials (fEPSPs) before and after cFC. After cFC, the commissural pathway’s synaptic strength was potentiated. Although recordings were conducted during the wake phase of the light/dark cycle, the mice slept more in the post-conditioning period than in the pre-conditioning period. Relative to awake periods, in non-rapid eye movement (NREM) sleep the fEPSPs were larger in both pre- and post-conditioning periods. Therefore, to avoid sleep-related fEPSP changes confounding the effects of cFC learning, we accounted for the effects of sleep and still found long-term potentiation of the commissural pathway, but no significant long-term depression. Synaptic strength changes were not found in the no-shock, control group that simply explored the fear-conditioning chamber, indicating that exploration of the novel place did not produce the measurable effects caused by cFC. These results show that following cFC, the CA1 commissural pathway is potentiated, likely contributing to the functional integration of the left and right hippocampi in fear memory consolidation. In addition, the cFC paradigm produces significant changes in an animal’s behavioral state, which are observable as proximal changes in sleep patterns.

A 28 Day Clinical Assessment of a Lactic Acid-containing Antimicrobial Intimate Gel Wash Formulation on Skin Tolerance and Impact on the Vulvar Microbiome

While intimate feminine hygiene products are widely used as part of daily cleansing routines, little is known about how these products impact the vulvovaginal area and its microbiome stability. This 4 week clinical study assessed tolerance of a novel gel wash containing lactic acid (pH 4.2) for external daily use when used on the external genital area and its effects on skin moisturization, vulvar skin pH, and the vulvar microbiome. After a 7 day pre-study conditioning period, 36 healthy females in three balanced age groups (18–29, 30–44, and 45–55 years) used the gel wash to cleanse their external genital area (mons pubis and vulva) and entire body at least once per day for 28 days. Skin tolerance of the gel wash was assessed by the gynecologist. Effects of the gel wash on vulvar skin microbiota were studied by performing bacterial 16S rRNA and fungal internal transcribed spacer (ITS) microbial richness and diversity analysis. Based on gynecologic assessment after 28 days of use, the gel wash showed acceptable tolerance, with no signs of increased dryness, redness, edema, itching, stinging, or burning. Use of the gel wash was associated with a significant increase in both short-term (single application) and longer-term (daily use for 28 days) skin moisturization. There was no significant change in vulvar skin pH over time with daily product use, and the gel wash did not significantly affect the natural vulvar microbiome species richness or diversity for bacteria or fungi. Results showed that this gel wash is a mild, moisturizing cleanser that maintains the natural pH and microbial diversity of vulvar skin. To our knowledge, this was the first study to assess the effect of an antimicrobial feminine gel wash on the natural pH and vulvar microbiome habitat of the skin using bacterial 16S rRNA and fungal ITS genetic sequencing techniques.

Assessing the affective component of pain, and the efficacy of pain control, using conditioned place aversion in calves

Pain in animals is typically assessed using reflexive and physiological responses. These measures allow inferences regarding nociception but provide little basis for conclusions about the affective component of pain (i.e. how negatively the experience is perceived). Calves routinely undergo painful procedures on commercial farms, including hot-iron disbudding, providing a convenient model to study pain in animals. The aim of this study was to investigate the affective component of post-procedural pain due to hot-iron disbudding, using conditioned place aversion. Calves ( n = 31) were subjected to two procedures (one bud at a time): one without post-procedural pain control and the other with the use of a nonsteroidal anti-inflammatory drug (either meloxicam ( n = 16) or ketoprofen ( n = 15)). All procedures included the use of local anaesthesia (lidocaine). Place conditioning was tested 2 days after the last treatment by allowing calves to freely roam between the pens where they had previously been disbudded. Calves spent more time, and lay down more frequently, in the pen where they received meloxicam compared with the pen where they only received a local block. Surprisingly, calves avoided the pen where they received ketoprofen compared with the control treatment pen. We hypothesize that the shorter duration of action of ketoprofen resulted in increasing pain at the end of the conditioning period, explaining the increased aversion to this treatment. These results illustrate the value of place conditioning paradigms to assess the affective component of pain in animals, and suggest that the animal's evaluation of painful events depends upon the time course of when the pain is experienced.

Maze learning and memory in a decapod crustacean

Spatial learning is an ecologically important trait well studied in vertebrates and a few invertebrates yet poorly understood in crustaceans. We investigated the ability of European shore crabs, Carcinus maenas , to learn a complex maze over four consecutive weeks using food as a motivator. Crabs showed steady improvement during this conditioning period in both the time taken to find the food and in the number of wrong turns taken. Crabs also clearly remembered the maze as when returned two weeks later but without any food, they all returned to the end of the maze in under 8 min. Crabs that had not been conditioned to the maze (naive animals) took far longer to reach the end, and many (42%) did not venture to the end of the maze at all during the 1 h study period. This study provides an initial description of spatial learning in a benthic decapod; a better appreciation of this adaptive trait in these animals will develop our understanding of resource exploitation by benthic crustaceans and their ecological roles.

Time Development of Voltage Frequency Dependence of Partial Discharge Activity in Voids

Condition assessment of high voltage equipment based on partial discharge measurements is often performed after a voltage pre-conditioning period. The aim of this paper is to present results from experimental examinations of time variance of partial discharge activity and to propose physical explanations of the phenomena observed. Experiments were performed on laboratory made 3 mm thick discs of generator bar insulation, consisting of mica and glass fiber reinforced epoxy with a 0.5 mm thick cylindrical void surfaces of 10 mm in diameter. The effect of conducting and insulating void surfaces was examined using copper tape as upper and lower electrodes of the voids. All objects were tested by 12 one-minute long AC voltage frequency sweeps at frequencies from 50 Hz to 0.1 Hz distributed in time from start of the experiment, after initial one-hour constant 50 Hz voltage application and during object short-circuiting for 20 h. The main result shows that in case of insulating voids the apparent charges vanished after the one-hour constant 50 Hz voltage application. After a grounding period of 5 minutes, the charge magnitudes slowly increased with time until reaching steady state after about 4-8 hours. Test objects with conductive void surfaces showed such reduction in case of PD testing at 0.1 Hz only. At voltage frequencies above 10 Hz the measured PD magnitudes were found to be nearly constant, close to the expected high theoretical value. This indicate that PD by-products strongly affected the void resistivity and thereby affect the PD activity of insulating voids. These by-products are temporary and disappear with time.