-
Animals : An Open Access Journal From... Oct 2023Refinement of experimental procedures in animal research has the objective of preventing and minimizing pain/distress in animals, including the euthanasia period. This...
Refinement of experimental procedures in animal research has the objective of preventing and minimizing pain/distress in animals, including the euthanasia period. This study aimed to evaluate pain associated with six methods of euthanasia in Wistar rats (injectable, inhalational, and physical), by applying the Rat Grimace Scale (RGS), comparing the scores, and determining the method with the highest score that might indicate pain for laboratory rodents. Sixty adult male and female Wistar rats were used and assigned to six treatments: pentobarbital, CO, decapitation, isoflurane, ketamine + xylazine, and ketamine + CO. Video recording to assess the RGS scores was performed in four events: basal: 24 h before the procedure; Ti: three minutes before the procedure; Ti: during the application of the euthanasia method; and Ti: immediately after the application until LORR. The main findings of this study showed that, during Ti, decapitation and ketamine + xylazine had the highest scores (0.6 ± 0.26 and 0.6 ± 0.16, respectively) ( < 0.0001), while at Ti, CO (0.9 ± 0.18) and isoflurane (1.2 ± 0.20) recorded the highest scores ( < 0.0001). According to the present results, decapitation and ketamine + xylazine elicited short-term acute pain, possibly due to tissue damage caused by both methods (injection and guillotine). In contrast, isoflurane's RGS scores recorded during Ti might be associated with nociception/pain due to the pungency of the drug or to the pharmacological muscle relaxant effect of isoflurane. Further research is needed to establish a comprehensive study of pain during euthanasia, where RGS could be used minding the limitations that anesthetics might have on facial expression.
PubMed: 37893885
DOI: 10.3390/ani13203161 -
Medicina (Kaunas, Lithuania) Oct 2023: Periodontitis is an inflammatory disease in the supporting tissues of the teeth caused by specific microorganisms or groups of microorganisms. bacterium is the...
: Periodontitis is an inflammatory disease in the supporting tissues of the teeth caused by specific microorganisms or groups of microorganisms. bacterium is the keystone pathogen in periodontitis, so even at low concentrations, it has a considerable influence on the oral community. Antimicrobials and antiplaque agents can be used as adjunctive therapy for periodontitis treatment. Konjac glucomannan (KGM), as a natural polysaccharide, has flavonoid (3,5-diacetyltambulin) and triterpenoids (ambylon) compounds that show antibacterial activity. This research aims to analyze the antibacterial activity of KGM on animal and in vitro periodontitis models. : The animal study divided 48 mice into four groups (control, KGM, periodontitis, KGM + periodontitis). Mice were given an intervention substance by oral gavage from day 1 to day 14, periodontitis was induced on day 7, and decapitation was performed on day 14. Samples from the right maxillary jaw of mice were used for histological preparations and morphometrics analysis. In vitro studies were carried out by adding several concentrations of KGM (25, 50, and 100 μg/mL) into a planktonic and biofilm. : In the animal model, KGM could prevent alveolar bone loss in the periodontitis mice model, both in histologic and morphometrics assessments. In vitro, KGM had antibacterial activity against with better bacteriostatic (15-23%) than bactericidal (11-20%) ability, proven by its ability to inhibit proliferation. : KGM can be considered to have the potential as an antibacterial agent to prevent periodontitis. The prevention of periodontitis may improve patient well-being and human quality of life.
Topics: Humans; Animals; Mice; Quality of Life; Periodontitis; Disease Models, Animal; Anti-Bacterial Agents
PubMed: 37893496
DOI: 10.3390/medicina59101778 -
Fa Yi Xue Za Zhi Aug 2023To explore the potential biomarkers for the diagnosis of primary brain stem injury (PBSI) by using metabonomics method to observe the changes of metabolites in rats with...
OBJECTIVES
To explore the potential biomarkers for the diagnosis of primary brain stem injury (PBSI) by using metabonomics method to observe the changes of metabolites in rats with PBSI caused death.
METHODS
PBSI, non-brain stem brain injury and decapitation rat models were established, and metabolic maps of brain stem were obtained by LC-MS metabonomics method and annotated to the HMDB database. Partial least square-discriminant analysis (PLS-DA) and random forest methods were used to screen potential biomarkers associated with PBSI diagnosis.
RESULTS
Eighty-six potential metabolic markers associated with PBSI were screened by PLS-DA. They were modeled and predicted by random forest algorithm with an accuracy rate of 83.3%. The 818 metabolic markers annotated to HMDB database were used for random forest modeling and prediction, and the accuracy rate was 88.9%. According to the importance in the identification of cause of death, the most important metabolic markers that were significantly up-regulated in PBSI group were HMDB0038126 (genipinic acid, GA), HMDB0013272 (-lauroylglycine), HMDB0005199 [()-salsolinol] and HMDB0013645 (,-dimethylsphingosine).
CONCLUSIONS
GA, -lauroylglycine, ()-salsolinol and ,-dimethylsphingosine are expected to be important metabolite indicators in the diagnosis of PBSI caused death, thus providing clues for forensic medicine practice.
Topics: Rats; Animals; Metabolomics; Brain Injuries; Biomarkers; Brain Stem
PubMed: 37859476
DOI: 10.12116/j.issn.1004-5619.2022.420510 -
International Immunopharmacology Nov 2023The therapeutic effect of liposomal IL-22 versus non-liposomal IL-22 on liver fibrosis was investigated. IL-22 (5 µg/ml) was incorporated into negative charged...
The therapeutic effect of liposomal IL-22 versus non-liposomal IL-22 on liver fibrosis was investigated. IL-22 (5 µg/ml) was incorporated into negative charged liposomes. Schistosoma mansoni infected mice were treated with liposomal IL-22 for either 7 or 14 days before decapitation. Liver and spleen were removed and splenocytes were isolated for in vitro investigations. TNF-α, IL-17, IL-22 and IgE levels were assessed. Hepatic granulomas were counted, granuloma index and its developmental stages were calculated. Hepatic expressions of STAT3, β-catenin and let-7a miRNA were evaluated. Liposomal IL-22 size was clustered around 425.9 ± 58.0 nm with negative zeta potential (-18.8 ± 1.3 mV). After 14 days, 65.5% of IL-22 was released from liposomal IL-22 as was gradually observed in vitro. Liposomal IL-22 significantly (p < 0.05) decreased IL-17 level (-33.1%) of healthy splenocytes compared to non-liposomal IL-22. In vivo therapeutic effect of liposomal IL-22 revealed a significant (p < 0.05) decrease in hepatic granuloma index (-22.1%) and levels of TNF-α (-49.2%) and IL-17 (-57.3%), but a marked increase in IL-22 (64.2%) and IgE (196.1%) levels comparing to non-liposomal IL-22. Three developmental stages of hepatic granuloma (NE, EP, and P) were observed in liposomal and non-liposomal IL-22 groups (79.6 ± 1.7 and 81.8 ± 8.7, respectively, P < 0.05), with higher relative frequency of EP stage. Additionally, liposomal IL-22 treatment increased hepatic expression of STAT3 (21.7 fold change) and let-7a (3.6 fold change) and reduced β-catenin expression (0.6 fold change) compared to healthy mice. Conclusively, liposomal IL-22 seems more effective in the treatment of liver fibrosis resulting from S. mansoni infection than non-liposomal IL-22.
Topics: Mice; Animals; Interleukin-17; beta Catenin; Tumor Necrosis Factor-alpha; Liposomes; Liver Cirrhosis; Liver; MicroRNAs; Granuloma; Immunoglobulin E; Interleukin-22
PubMed: 37827055
DOI: 10.1016/j.intimp.2023.111015 -
European Review For Medical and... Sep 2023The objective of our study was to evaluate whether ovarian suppression by two different hormonal methods may spare the ovary the cytotoxic effects of isotretinoin in a...
The protective effects of hormonal suppression by a gonadotropin-releasing hormone agonist or an oral contraceptive on the decreased ovarian reserve in female rats exposed to isotretinoin.
OBJECTIVE
The objective of our study was to evaluate whether ovarian suppression by two different hormonal methods may spare the ovary the cytotoxic effects of isotretinoin in a rat model.
MATERIALS AND METHODS
Four groups (n=8 Sprague-Dawley albino rats per group) were studied: control (Group I), 7.5 mg/kg/day isotretinoin (Group II), isotretinoin plus the combination of 0.030 mg ethinyl estradiol/0.15 mg levonorgestrel (combined oral contraceptive, COC), and isotretinoin plus 100 μg (microgram) leuprolide acetate (GnRHa) (Group III and IV, respectively). Four rats from each group were decapitated on the 30th day of treatment, and the remaining rats were decapitated on the 30th day of untreated follow-up. Serum anti-Mullerian hormone (AMH) concentrations, healthy and atretic follicle numbers, and apoptotic activity of follicles in oophorectomy specimens were compared between the groups.
RESULTS
There were no significant differences in AMH levels among the study groups before, immediately after (first month), and one month after their last medication (second month) (p=0.08, 0.47, and 0.08, respectively). At the end of the first month, the control group had a higher median count of healthy primordial follicles compared to the study groups: 13.5 (8-22), 5.5 (3-11), 6 (2-13), and 1 (0-1) in control, isotretinoin, isotretinoin+COC, and isotretinoin+GnRHa groups, respectively (p=0.02). However, there was no statistically significant difference in the number of healthy primordial follicles between the groups one month after the last medication (p=0.33). The median atretic antral follicle counts in the first month were 2 (1-4), 3.5 (1-4), 0 (0-2), and 0 (0-0) in the control, isotretinoin, isotretinoin+COC, and isotretinoin+GnRHa groups, respectively (p=0.02). Otherwise, there were no significant differences in other types of follicles among the control and treated groups (p>0.05). There was also no statistical difference between the groups regarding immunostaining intensity for active caspase-3 evaluated in the first or second month of treatment (p=0.8 and 0.2, respectively).
CONCLUSIONS
Our results show that GnRH agonists or COC have no protective effects on ovarian reserve when co-administered with isotretinoin in the rat model.
Topics: Female; Rats; Animals; Humans; Contraceptives, Oral; Isotretinoin; Ovarian Reserve; Rats, Sprague-Dawley; Anti-Mullerian Hormone; Immunologic Factors; Gonadotropin-Releasing Hormone
PubMed: 37782196
DOI: 10.26355/eurrev_202309_33808 -
Frontiers in Plant Science 2023Sweet sorghum has emerged as a promising source of bioenergy mainly due to its high biomass and high soluble sugar yield in stems. Studies have shown that...
Sweet sorghum has emerged as a promising source of bioenergy mainly due to its high biomass and high soluble sugar yield in stems. Studies have shown that loss-of-function locus alleles have been selected during sweet sorghum domestication, and decapitation can further boost sugar accumulation in sweet sorghum, indicating that the potential for improving sugar yields is yet to be fully realized. To maximize sugar accumulation, it is essential to gain a better understanding of the mechanism underlying the massive accumulation of soluble sugars in sweet sorghum stems in addition to the locus. We performed a transcriptomic analysis upon decapitation of near-isogenic lines for mutant (, juicy stems, and green leaf midrib) and functional (, dry stems and white leaf midrib) alleles at the locus. Our analysis revealed that decapitation suppressed photosynthesis in leaves, but accelerated starch metabolic processes in stems. negatively correlates with sugar levels supported by genotypes ( vs. ), treatments (control vs. decapitation), and developmental stages post anthesis (3d vs.10d). locus gene and other programmed cell death-related genes were downregulated by decapitation, while sugar transporter-encoding gene was induced. Both and were detected in phloem companion cells by RNA assay. Loss of the homolog, in led to a sugar accumulation increase. This study provides new insights into sugar accumulation enhancement in bioenergy crops, which can be potentially achieved by reducing reproductive sink strength and enhancing phloem unloading.
PubMed: 37767289
DOI: 10.3389/fpls.2023.1233813 -
Animals : An Open Access Journal From... Sep 2023Refinement is one of the principles aiming to promote welfare in research animals. The techniques used during an experimental protocol, including euthanasia selection,...
Refinement is one of the principles aiming to promote welfare in research animals. The techniques used during an experimental protocol, including euthanasia selection, must prevent and minimize suffering. Although the current euthanasia methods applied to laboratory rodents are accepted, the controversial findings regarding the potential stress/distress they can cause is a field of research. The objective was to assess the thermal response of Wistar rats during various euthanasia methods using infrared thermography (IRT) to determine the method that prevents or diminishes the stress response and prolonged suffering. Pentobarbital (G), CO (G), decapitation (G), isoflurane (G), ketamine + xylazine (G), and ketamine + CO (G) were evaluated at five evaluation times with IRT to identify changes in the surface temperature of four anatomical regions: ocular (T°), auricular (T°), interscapular (T°), and caudal (T°). Significant differences ( < 0.05) were found in G and G, registering temperature increases from the administration of the drug to the cessation of respiratory rate and heart rate. Particularly, isoflurane showed a marked thermal response in T°, T°, T°, and T°, suggesting that, in general, inhalant euthanasia methods induce stress in rats and that isoflurane might potentially cause distress, an effect that must be considered when deciding humane euthanasia methods in laboratory rodents.
PubMed: 37760220
DOI: 10.3390/ani13182820 -
Journal of Plant Research Nov 2023Plants are exposed to a variety of biotic and abiotic stresses, including wounding at the stem. The healing process (tissue reunion) begins immediately after stem...
Plasmodesmata callose binding protein 2 contributes to the regulation of cambium/phloem formation and auxin response during the tissue reunion process in incised Arabidopsis stem.
Plants are exposed to a variety of biotic and abiotic stresses, including wounding at the stem. The healing process (tissue reunion) begins immediately after stem wounding. The plant hormone auxin plays an important role during tissue reunion. In decapitated stems, auxin transport from the shoot apex is reduced and tissue reunion does not occur but is restored by application of indole-3-acetic acid (IAA). In this study, we found that plasmodesmata callose binding protein 2 (PDCB2) affects the expansion of the cambium/phloem region via changes in auxin response during the process of tissue reunion. PDCB2 was expressed in the cortex and endodermis on the incised side of stems 1-3 days after incision. PDCB2-knockout plants showed reduced callose deposition at plasmodesmata and DR5::GUS activity in the endodermis/cortex in the upper region of the incision accompanied by an increase in size of the cambium/phloem region during tissue reunion. In addition, PIN(PIN-FORMED)3, which is involved in lateral auxin transport, was induced by auxin in the cambium/phloem and endodermis/cortex in the upper part of the incision in wild type, but its expression of PIN3 was decreased in pdcb2 mutant. Our results suggest that PDCB2 contributes to the regulation of cambium/phloem development via auxin response.
Topics: Arabidopsis; Phloem; Cambium; Arabidopsis Proteins; Carrier Proteins; Plasmodesmata; Indoleacetic Acids; Gene Expression Regulation, Plant
PubMed: 37707645
DOI: 10.1007/s10265-023-01494-0 -
Ultrasound in Obstetrics & Gynecology :... Feb 2024
Topics: Humans; Female; Decapitation; Ultrasonography; Vagina
PubMed: 37698521
DOI: 10.1002/uog.27467 -
Journal of Molecular Recognition : JMR Nov 2023One of the leading causes of acute lung injury, which is linked to a high death rate, is pulmonary fat embolism. Increases in proinflammatory cytokines and the...
One of the leading causes of acute lung injury, which is linked to a high death rate, is pulmonary fat embolism. Increases in proinflammatory cytokines and the production of free radicals are related to the pathophysiology of acute lung injury. Antioxidants that scavenge free radicals play a protective role against acute lung injury. Gossypin has been proven to have antioxidant, antimicrobial, and anti-inflammatory properties. In this study, we compared the role of Gossypin with the therapeutically used drug Dexamethasone in the acute lung injury model caused by oleic acid in rats. Thirty rats were divided into five groups; Sham, Oleic acid model, Oleic acid+Dexamethasone (0.1 mg/kg), Oleic acid+Gossypin (10 and 20 mg/kg). Two hours after pretreatment with Dexamethasone or Gossypin, the acute lung injury model was created by injecting 1 g/kg oleic acid into the femoral vein. Three hours following the oleic acid injection, rats were decapitated. Lung tissues were extracted for histological, immunohistochemical, biochemical, PCR, and SEM imaging assessment. The oleic acid injection caused an increase in lipid peroxidation and catalase activity, pathological changes in lung tissue, decreased superoxide dismutase activity, and glutathione level, and increased TNF-α, IL-1β, IL-6, and IL-8 expression. However, these changes were attenuated after treatment with Gossypin and Dexamethasone. By reducing the expression of proinflammatory cytokines and attenuating oxidative stress, Gossypin pretreatment provides a new target that is equally effective as dexamethasone in the treatment of oleic acid-induced acute lung injury.
PubMed: 37696682
DOI: 10.1002/jmr.3058