Search Results (24)

Background: This study aimed to develop a model for understanding stress-induced sleep disturbances and to explore the potential interactions between sleep disturbances and mood disturbances. Methods: The chronic unpredictable mild stress (CUMS) group was established using the CUMS method, while the CUMS+Noise group was subjected to an additional 8-h exposure to noise in conjunction with the CUMS protocol. Each group was tested for anxiety and depressive-like behavior using the open-field, elevated plus maze, tail suspension, and forced swimming tests in male C57BL/6J mice. Subsequently, we assessed sleep status using sleep recordings and a standardized scoring system alongside the pentobarbital sodium-induced sleep test. Results: The mice in both model groups exhibited anxiety-like behavior. Sleep disturbances observed in the CUMS+Noise group were characterized by disruptions in sleep duration and circadian rhythm. This observation was supported by a marked reduction in multiple sleep time intervals and single sleep duration, as well as a significant increase in sleep duration at the final time interval of ZT23-24. To further investigate the potential mechanisms of interaction, we conducted an analysis of hub genes present in the hippocampal sequencing data utilizing weighted gene co-expression network analysis (WGCNA). Pearson correlation analysis revealed a significant association between the hub genes Alb, P2rx1, and Npsr1 and key phenotypic traits. However, PCR experiments indicated that only Alb showed a significant difference, which aligns with the sequencing results. Conclusions: Albumin is a crucial transporter protein for thyroid hormones and plays a vital role in their metabolism. The interaction between sleep disorders and anxiety-like behavior may be closely linked to the dysfunctional transportation of thyroid hormones by albumin. Full article

Ziziphi Spinosae Semen saponins (ZSSS) show sedative–hypnotic activity but have very low bioavailability, potentially due to their conversion into bioactive metabolites by gut microbiota. In this study, the biotransformation of ZSSS by gut microbiota from healthy humans and patients with insomnia in vitro was analyzed. A total of 21 prototype compounds and 49 metabolites were identified using UHPLC-Q-Orbitrap-MS. Deglycosylation, deoxygenation, dehydration, and deacylation were detected in both healthy individuals and insomniacs. However, oxidation and hydrogenation were uniquely observed in insomniacs. ZSSS can enhance beneficial bacteria, such as Veillonella, Dialister, and Bacteroides. ZSSS can promote the synthesis of short-chain fatty acids (SCFAs), especially acetic acid, propionic acid, and butyric acid. Furthermore, it was found that the sedative–hypnotic activity of ZSSS was enhanced after biotransformation, as determined by a sodium pentobarbital-induced sleeping test (SPST), open-field behavior test (OFBT), and molecular docking experiment (MDE). These results collectively offer valuable insight into the mechanism of action of ZSSS. Full article

Insomnia seriously affects people’s health and daily life. There is a growing interest in sleep-promoting agents from natural sources. Nelumbinis Plumula (NP), a traditional Chinese medicine with dual food-medicine homology, has the effects of clearing the heart and calming the mind, showing promising efficacy in treating insomnia. In this study, the effects of NP extract, total alkaloid extract of NP, and crude polysaccharide of NP were measured in para-chlorophenylalanine-induced insomnia rats combined with the pentobarbital sodium experiment. The results indicated both total alkaloid extract and NP total extract could improve insomnia in rats, with the total alkaloid extract demonstrating a stronger effect than NP total extract. Total alkaloid extract significantly prolonged sleep duration and shortened sleep latency. Therefore, total alkaloids in NP appeared to be the main pharmacological substances that exerted sedative effect. Simultaneously, total alkaloid extract could increase the GABA level and reduce the DA level as well as affect the activities of GABRA1, DRD2, 5-HT1A, and AChE proteins. This study can lay an experimental foundation for the further development and application of NP as a remedy for treating insomnia. Full article

Diabetic nephropathy (DN) is a serious complication of diabetes mellitus; oxidative stress plays a key role in the pathogenesis of DN. The objective of this study was to evaluate the antioxidant effect of vitamin E on diabetic nephropathy. A control group and three groups of rats with streptozotocin-induced diabetes mellitus (untreated diabetic rats and diabetic rats treated with vitamin E 250 and 500 mg/kg) were studied. After 4 weeks of treatment, the kidneys were removed under anesthesia with sodium pentobarbital. The kidneys were weighed, the AT₁ and AT₂ receptor expression was measured by Western blot, and the activities of glutathione peroxidase, catalase, and superoxide dismutase were determined in the renal cortex. Rats with diabetes mellitus had hyperglycemia, increased food and water consumption, and higher urinary volume than control rats. In diabetic rats (DM), kidney hypertrophy was observed and measured by kidney weight, protein/DNA ratio in the renal cortex, and proximal tubular cell area; proteinuria and reduced creatinine clearance were observed. AT₁ and AT₂ receptor expression in the kidney cortex of DM rats increased significantly compared to normoglycemic rats; antioxidant enzyme activities were decreased; treatment with vitamin E reversed kidney hypertrophy and reduced proteinuria; reduction in expression of AT₁ and AT₂ receptors was associated with increased antioxidant activity. Thus, treatment with vitamin E slows the progress of DN. Full article

Background and Objective: The alteration of mitochondrial functions, especially the opening of the mitochondrial permeability transition pore (mPTP), has been proposed as a key mechanism in the development of lesions in cerebral ischemia, wherefore it is considered as an important target for drugs against ischemic injury. In this study, we aimed to investigate the effects of mitochondrial complex I inhibitors as possible regulators of mPTP using an in vitro brain ischemia model of the pentobarbital/ketamine (PBK)-anesthetized rats. Results: We found that PBK anesthesia itself delayed Ca²⁺-induced mPTP opening and partially recovered the respiratory functions of mitochondria, isolated from rat brain cortex and cerebellum. In addition, PBK reduced cell death in rat brain slices of cerebral cortex and cerebellum. PBK inhibited the adenosine diphosphate (ADP)-stimulated respiration of isolated cortical and cerebellar mitochondria respiring with complex I-dependent substrates pyruvate and malate. Moreover, pentobarbital alone directly increased the resistance of isolated cortex mitochondria to Ca²⁺-induced activation of mPTP and inhibited complex I-dependent respiration and mitochondrial complex I activity. In contrast, ketamine had no direct effect on functions of isolated normal cortex and cerebellum mitochondria. Conclusions: Altogether, this suggests that modulation of mitochondrial complex I activity by pentobarbital during PBK anesthesia may increase the resistance of mitochondria to mPTP opening, which is considered the key event in brain cell necrosis during ischemia. Full article

Daytime dysfunction, including symptoms like sleepiness, poor memory, and reduced responsiveness, is not well researched. It is crucial to develop animal models and study the biological mechanisms involved. We simulated sleep disorders through sleep deprivation, and stressful stimuli were used to establish daytime functional animal models. We used tests like the sodium pentobarbital sleep synergy test and the DSI telemetry system to measure sleep duration and structure. We also used tests like the Morris water maze, open field test, grip test, and baton twirling test to assess mental and physical fatigue. To assess the intrinsic biological mechanisms, we measured sleep–wake-related neurotransmitters and related receptor proteins, circadian rhythm-related proteins and cognition-related proteins in hypothalamus tissue, and oxidative stress, inflammatory factors, S100β, and HPA axis-related indexes in serum. Multi-factor sleep deprivation resulted in the disruption of sleep–wakefulness structure, memory–cognitive function degradation, decreased grip coordination, and other manifestations of decreased energetic and physical strength. The intrinsic biological mechanisms were related to the disturbed expression of sleep–wake, circadian rhythm, memory–cognition-related proteins, as well as the significant elevation of inflammatory factors, oxidative stress, the HPA axis, and other related indicators. Intrinsically related biological mechanisms and reduced sirt1 expression can lead to disruption of circadian rhythms; resulting in disruption of their sleep–wake-related neurotransmitter content and receptor expression. Meanwhile, the reduced expression of sirt1 also resulted in reduced expression of synapse-associated proteins. This study prepared an animal model of daytime dysfunction by means of multi-factor sleep deprivation. With sirt1 as a core target, the relevant biological mechanisms of neurological disorders were modulated. Full article

This study was designed to determine how veterinarians define a good euthanasia experience. This information is used to generate a working definition of companion animal euthanasia that aligns with animal welfare standards and pet owners’ expectations. An electronic survey distributed via veterinary-related social media (Facebook, Instagram) and listservs were completed by 249 veterinarians who perform feline and/or canine euthanasia. Our results suggest that very few veterinarians feel their veterinary school training adequately prepared them for euthanasia. When veterinarians were asked to rank a list of physiologic conditions and anatomical traits in order of euthanasia-related concerns, respiratory distress was ranked the highest, while the most concerning physical changes were reported to be indications or impressions of seizures or pain. The most commonly reported euthanasia injection technique performed by participants was intravenous administration of pentobarbital sodium (97%), and most veterinarians preferred having owners present (57%) or having no preference (38%) during euthanasia. Results suggest that veterinarians want a pain-free, anxiety-free experience for the patient, appreciate the use of sedatives before euthanasia, and feel that when available and appropriate, home euthanasia offers several benefits. This understanding of the numerous aspects involved in a good euthanasia experience can help inform the creation of an updated definition of companion animal euthanasia that strives to prioritize the welfare of the patient as well as the needs and expectations of the pet owner. Full article

Pentobarbital is a drug of choice to limit motion in children during paediatric procedural sedations (PPSs). However, despite the rectal route being preferred for infants and children, no pentobarbital suppositories are marketed, and therefore they must be prepared by compounding pharmacies. In this study, two suppository formulations of 30, 40, 50, and 60 mg of pentobarbital sodium were developed using hard-fat Witepsol^® W25 either alone (formulation F1) or with oleic acid (formulation F2). The two formulations were subjected to the following tests described in the European Pharmacopoeia: uniformity of dosage units, softening time, resistance to rupture, and disintegration time. The stability of both formulations was also investigated for 41 weeks of storage at 5 ± 3 °C using a stability-indicating liquid chromatography method to quantify pentobarbital sodium and research breakdown product (BP). Although both formulae were compliant to uniformity of dosage, the results were in favour of a faster disintegration of F2 compared to F1 (−63%). On the other hand, F1 was found to be stable after 41 weeks of storage unlike F2 for which several new peaks were detected during the chromatographic analysis, suggesting a shorter stability of only 28 weeks. Both formulae still need to be clinically investigated to confirm their safety and efficiency for PPS. Full article

Lobeline is an alkaloid derived from the leaves of an Indian tobacco plant (Lobelia inflata), which has been prepared by chemical synthesis. It is classified as a partial nicotinic agonist and has a long history of therapeutic usage ranging from emetic and respiratory stimulant to tobacco smoking cessation agent. The presence of both cis and trans isomers in lobeline is well known, and many studies on the relationship between the structure and pharmacological activity of lobeline and its analogs have been reported. However, it is a remarkable fact that no studies have reported the differences in pharmacological activities between the two isomers. In this article, we found that different degrees of isomerization of lobeline injection have significant differences in respiratory excitatory effects in pentobarbital sodium anesthetized rats. Compared with cis-lobeline injections, the respiratory excitatory effect was significantly reduced by 50.2% after administration of injections which contained 36.9% trans-lobeline. The study on the influencing factors of isomerization between two isomers shown that this isomerization was a one-way isomerism and only converted from cis to trans, where temperature was the catalytic factor and pH was the key factor. This study reports a new discovery. Despite the widespread use of ventilators, first-aid medicines such as nikethamide and lobeline has retired to second line, but as a nonselective antagonist with high affinity for a4b2 and a3b2 nicotinic acetylcholine receptors (nAChRs). In recent years, lobeline has shown great promise as a therapeutic drug for mental addiction and nervous system disorders, such as depression, Alzheimer disease and Parkinson disease. Therefore, we suggest that the differences between two isomers should be concerned in subsequent research papers and applications. Full article

Background: The purpose of this study was to measure the preventive and regenerative effect of glutamine and probiotics induced by alcohol injury in Holtzman rats. Methods: Analytical, experimental and prospective study. The population consisted of 56 male rats between 300 and 350 g, distributed in three experimental phases: Pre-pilot phase PPP (6 rats), Pilot phase PP (10 rats), and Experimental phase EP (40 rats). In the pilot phase, 10 rats were subjected to damage with 8.5% ethanol, which was given intragastrically. The dosage was calculated for 10 rats in two groups: the first with 7.5 mL/kg in 5 rats and the second with 8.5 mL/kg in 5 rats. The experimental phase was performed in 40 rats divided into 6 groups, the negative control group (healthy), positive control group (injured), preventive experimental group (glutamine and glutamine with probiotic) and regenerative experimental group (glutamine and glutamine with probiotic). At the end of each phase, the rats were sacrificed with sodium pentobarbital (Halathal) and a portion of their stomachs was stored in formol. Results: The evaluation of stomach tissue samples (desquamation, erythema, hyperemia) showed that in the preventive phase, glutamine shows effectiveness in comparison to glutamine with probiotic. In the regenerative phase, glutamine and glutamine with probiotic did not show significant differences. Conclusions: Glutamine and probiotics can potentially serve as a therapy for the treatment for gastritis. Full article

In 2021, a shortage in the supply of the euthanasia drug pentobarbital sodium affected animal care professionals around the world, including in the United States and Canada. Pentobarbital sodium is the drug of choice for companion animal euthanasia in both countries. The decreased availability of pentobarbital sodium affected a number of animal care industries, forcing conservation of the drug and the use of alternative methods and other agents to facilitate humane death for all manner of animal species. Veterinary medical groups, laboratory research institutions, and the animal sheltering industry worked together to identify the best path forward to maintain routine euthanasia practices and to protect the welfare of animals. This article aims to explore the reasons behind the shortage and to highlight the necessary responses and adjustments made in order to continue providing euthanasia services in North America. Recommendations for handling future pentobarbital shortages are included. Full article

Drug addiction has become a worldwide problem, affecting millions of people across the globe. While the majority of mechanistic studies on drug addiction have been focused on the central nervous system, including the mesolimbic dopamine system, the peripheral actions of drugs of abuse remain largely unknown. Our preliminary study found that the systemic injection of cocaine increased peripheral skin temperature. This led us to our present study, which investigated the mechanisms underlying the increase in peripheral temperature following cocaine injection. Male Sprague Dawley rats were anesthetized with pentobarbital sodium, and peripheral skin temperature measurements were taken using a thermocouple needle microprobe and an infrared thermal camera. Cocaine injection caused an acute rise in peripheral body temperature, but not core body temperature, about 10 min after injection, and the temperature increases were occluded by systemic injection of dopamine D2 receptor antagonist L741,626, but not D1 receptor antagonist SCH23390. In addition, systemic administration of bromocriptine, a dopamine D2 receptor agonist, significantly increased peripheral temperature. Infrared thermal imaging showed that the thermal increases following cocaine injection were predominantly in the distal areas of the forelimbs and hindlimbs, relative to core body temperature. Treatment with cocaine or bromocriptine decreased the size of skin blood vessels without affecting the expression of dopamine D2 receptors. These results suggest that increased peripheral temperature in skin following cocaine injection is associated with the activation of the dopamine D2 receptor. Full article

Dengue, a mosquito-borne zoonotic disease, is the most common vector-borne disease in tropical and subtropical areas. In this study, we aim to demonstrate biological evidence of dengue virus infection in bats. A cross-sectional study was carried out in the departments of Cordoba and Sucre, Colombia. A total of 286 bats were captured following the ethical protocols of animal experimentation. The specimens were identified and euthanized using a pharmacological treatment with atropine, acepromazine and sodium pentobarbital. Duplicate samples of brain, heart, lung, spleen, liver, and kidney were collected with one set stored in Trizol and the other stored in 10% buffered formalin for histopathological and immunohistochemical analysis using polyclonal antibodies. Brain samples from lactating mice with an intracranial inoculation of DENV-2 were used as a positive control. As a negative control, lactating mouse brains without inoculation and bats brains negative for RT-PCR were included. Tissue sections from each specimen of bat without conjugate were used as staining control. In a specimen of Carollia perspicillata captured in Ayapel (Cordoba) and Phylostomus discolor captured in San Carlos (Cordoba), dengue virus was detected, and sequences were matched to DENV serotype 2. In bats RT-PCR positive for dengue, lesions compatible with viral infections, and the presence of antigens in tissues were observed. Molecular findings, pathological lesions, and detection of antigens in tissues could demonstrate viral DENV-2 replication and may correspond to natural infection in bats. Additional studies are needed to elucidate the exact role of these species in dengue epidemics. Full article

Stroke is a life-threatening condition that leads to the death of many people around the world. Reperfusion injury after ischemic stroke is a recurrent problem associated with various surgical procedures that involve the removal of blockages in the brain arteries. Lipid emulsion was recently shown to attenuate ischemic reperfusion injury in the heart and to protect the brain from excitotoxicity. However, investigations on the protective mechanisms of lipid emulsion against ischemia in the brain are still lacking. This study aimed to determine the neuroprotective effects of lipid emulsion in an in vivo rat model of ischemic reperfusion injury through middle cerebral artery occlusion (MCAO). Under sodium pentobarbital anesthesia, rats were subjected to MCAO surgery and were administered with lipid emulsion through intra-arterial injection during reperfusion. The experimental animals were assessed for neurological deficit wherein the brains were extracted at 24 h after reperfusion for triphenyltetrazolium chloride staining, immunoblotting and qPCR. Neuroprotection was found to be dosage-dependent and the rats treated with 20% lipid emulsion had significantly decreased infarction volumes and lower Bederson scores. Phosphorylation of Akt and glycogen synthase kinase 3-β (GSK3-β) were increased in the 20% lipid-emulsion treated group. The Wnt-associated signals showed a marked increase with a concomitant decrease in signals of inflammatory markers in the group treated with 20% lipid emulsion. The protective effects of lipid emulsion and survival-related expression of genes such as Akt, GSK-3β, Wnt1 and β-catenin were reversed by the intra-peritoneal administration of XAV939 through the inhibition of the Wnt/β-catenin signaling pathway. These results suggest that lipid emulsion has neuroprotective effects against ischemic reperfusion injury in the brain through the modulation of the Wnt signaling pathway and may provide potential insights for the development of therapeutic targets. Full article

To explore the joint toxicity and bio-accumulation of multi-heavy metals and potential chemoprevention strategies, Male Sprague Dawley (SD) rats (n = 30) were treated orally once a week for six months with 500mg/kg•bw of eight heavy metals which were commonly identified in aquatic products in the Ningbo area including chromium, manganese, nickel, copper, zinc, cadmium, mercury, and lead. At the same time, 200mg/kg•bw of epigallocatechin-3-gallate (EGCG), trisodium citrate dihydrate (TCD) or glutathione (GSH) were administered to evaluate their antagonistic effects against adverse effects of multi-heavy metal mixture. The Morris water maze test was used to evaluate spatial learning and memory in the treated rats. Then the rats were anesthetized by pentobarbital sodium (40 mg/kg•bw) to obtain blood samples for biochemical analysis and organs (heart, liver, spleen, lungs, kidneys, brain, testis) to be conducted for biopsy and organ coefficients. Inductively coupled plasma mass spectrometer (ICP-MS) was used to analyze the concentrations of heavy metals. Results indicated that six months of exposure to a multi-heavy metal mixture under this experimental dosage resulted in accumulation in organs and adverse effects on the blood, reproductive system, and liver function. EGCG, TCD or GSH all showed certain chemoprevention effects against the joint toxicity induced by the multi-heavy metal mixture and indicated alleviation and the potential mechanism that also included the promotion of excretion of metals to which animals were exposed. Full article