Experimental Animal Studies Research Articles

The effect of nitric oxide supplied to the oxygenator of the cardiopulmonary bypass on the functions and morphology of internal organs: experimental animal study

To evaluate the effect of NO added to the oxygenator during CPB on the liver and kidneys in pigs.Methods. The experiment was conducted on 10 pigs undergoing cardiac surgery using cardiopulmonary bypass (CPB). Animals of the experimental group (CPB-NO; n = 5) received NO with the gas mixture supplied to the oxygenator at 100 ppm. Animals of the control group (CPB-control; n = 5) did not receive NO via the oxygenator. The surgery lasted 4 hours and was followed by 12-hour postoperative monitoring. To assess the liver and kidney injury, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin, creatinine, neutrophil gelatinase-associated lipocalin (NGAL) were determined at baseline, at weaning from the CPB, 6 and 12 hours after weaning from the CPB. A pathomorphological study of the liver and kidneys was also performed.Results. A long period of CPB deliberately used in our experiment caused liver injury. In the CPB-control group, an increase in the ALT concentration was found: 43 (34; 44) U/l at baseline to 82 (53; 99) U/l 12 hours after CPB, p < 0.05. AST concentration in the CPB-control group increased from 25 (17; 26) U/l at baseline to 269 (164; 376) U/l 12 hours after CPB (p < 0.05). We found no significant increase in ALT and AST concentrations in the CPB-NO group. There were no significant differences in ALT and AST concentrations between CPB-NO and CPB-control groups at all study time points. In the CPB-control group, an increase in the creatinine level was found from 131 (129; 133) μmol/l at baseline to 273 (241; 306) μmol/l 12 hours after CPB (p < 0.05). We found no significant increase in creatinine levels in the CPB-NO group. Creatinine levels in the CPB-NO group were significantly lower than in the CPB-control group 12 h after weaning from CPB: 183 (168; 196) vs 273 (241; 306) μmol/l (p < 0.008). We found no significant differences between the study groups by the NGAL level. Morphology revealed characteristic alterative changes in tissues, circulatory disorders, and inflammatory infiltration of varying degrees of severity, which in combination is a manifestation of the acute damaging effect of CPB. In the CPB-NO group, the severity of pathological changes, measured in points, was lower than in the CPB-control group, but no statistically significant differences were found.Conclusion. NO added to the oxygenator gas mixture reduces liver and kidney injury during prolonged CPB. Further research is required.

Mutations in nonstructural proteins essential for pathogenicity in SARS-CoV-2-infected mice.

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) has resulted in substantial morbidity and mortality. The basis of severe disease in humans is difficult to determine without the use of experimental animal models. Mice are resistant to infection with ancestral strains of SARS-CoV-2, although many variants that arose later in the pandemic were able to directly infect mice. In almost all cases, viruses that naturally infected mice or were engineered to enable mouse infection required mouse passage to become virulent. In most cases, changes in structural and nonstructural changes occurred during mouse adaptation. However, the mechanism of increased virulence in mice is not understood. Here, using a recently described strain of mouse-adapted SARS-CoV-2 (rSARS2-MA30N501Y), we engineered a series of recombinant viruses that expressed a subset of the mutations present in rSARS2-MA30N501Y. Mutations were detected in the spike protein and in three nonstructural proteins (nsp4, nsp8, and nsp9). We found that infection of mice with recombinant SARS-CoV-2 expressing only the S protein mutations caused very mild infection. Addition of the mutations in nsp4 and nsp8 was required for complete virulence. Of note, all these recombinant viruses replicated equivalently in cultured cells. The innate immune response was delayed after infection with virulent compared to attenuated viruses. Further, using a lineage tracking system, we found that attenuated virus was highly inhibited in the ability to infect the parenchyma, but not the airway after infection. Together, these results indicate that mutations in both the S protein and nonstructural proteins are required for maximal virulence during mouse adaptation.IMPORTANCEUnderstanding the pathogenesis of coronavirus disease 2019 (COVID-19) requires the study of experimental animals after infection with severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2). For this purpose, several mouse-adapted SARS-CoV-2 strains have been developed. Here, using a strain of mouse-adapted virus that causes a range of diseases ranging from mild to severe, we show that mutations in both a structural protein [spike (S) protein] and nonstructural proteins are required for maximal virulence. Thus, changes in the S protein, the most widely studied viral protein, while required for mouse adaptation, are not sufficient to result in a virulent virus.

Effects of Quercetin Fraction from Moringa oleifera Leaf Extract on Oxidative Markers and Histological Profile of Carotid and Coronary Arteries: An Experimental Animal Study

Effects of Quercetin Fraction from Moringa oleifera Leaf Extract on Oxidative Markers and Histological Profile of Carotid and Coronary Arteries: An Experimental Animal Study

Bis-(2-Chloroisopropyl) ether.

Bis-(2-Chloroisopropyl) ether (BCIPE) was used as a solvent for fats, greases, paint, varnish removers, and in spotting and cleaning solutions. However, BCIPE has not been commercially manufactured or used for numerous years. In experimental animal studies, BCIPE is moderately toxic following acute oral, dermal, and inhalation routes of exposure. BCIPE is a severe eye irritant but not a dermal irritant or dermal sensitizer. BCIPE was not genotoxic or mutagenic in in vitro and in vivo assays; it was not toxic in a 3-generation reproductive dietary study in rats. Short-term, repeated inhalation and oral exposure in rats produced increased liver and kidney weights and congestion; dermal exposure in rabbits did not produce any observable adverse effects. BCIPE did not produce a statistically significant increase in tumors in two different 2-year dietary studies in mice and rats. In mice, technical grade BCIPE produced increased incidences of alveolar/bronchiolar adenomas in females, hepatocellular carcinomas in males, and a low incidence of forestomach hyperplasia (in both sexes at the high-dose). Further investigation with technical grade BCIPE concluded that these effects were species- and dose-specific with limited, if any, relevance to humans. The NOAEL of 400 ppm (15 mg/kg/day) from the 2-year dietary study in female rats was considered the point of departure for the health-based WEEL derivation. After adjustment for duration of exposure, interindividual variability, and intraindividual variability, an 8-h time-weighted average (TWA) WEEL value of 3 ppm (21 mg/m3) was derived. This exposure limit is expected to provide a significant margin of safety against any potential adverse health effects in workers.

Physiological occlusal force attenuates replacement root resorption of replanted teeth: an experimental animal study

BackgroundTooth avulsion represents the most severe form of dental trauma, necessitating tooth replantation as the primary treatment. However, the risk of replacement root resorption (RRR) poses a significant threat to tooth retention following replantation. This study preliminarily aimed to investigate the effect of physiological occlusal force on RRR after the replantation of avulsed teeth and to explore the potential underlying mechanisms.MethodsThirty-six 4-week-old male Sprague-Dawley rats underwent extraction and immediate replantation of their left maxillary molars. The rats were randomly divided into two major groups: the occluded (n = 18) group, where the opposite mandibular teeth were preserved; non-occluded (n = 18) group, where the opposite mandibular teeth were extracted. Within each major group, there were three subgroups corresponding to 7 days, 14 days, and 2 months, resulting in a total of six subgroups, (n = 6 per subgroup). The right maxillary first molars served as the normal control. Various periodontal characteristics were assessed using haematoxylin-eosin (H&E), tartrate-resistant acid phosphatase (TRAP) staining, and micro-computed tomography (micro-CT).ResultsHistological staining revealed that under occlusal force, the early stage (day 7) after tooth replantation mainly manifested as root surface resorption, especially in the non-occluded group, which gradually diminished over time. Cementum and periodontal ligament (PDL) repair was observed on day 14. Micro-CT analysis indicated a significant decrease in PDL width in the non-occluded group two months after replantation, consistent with the histological findings, signifying severe RRR in the non-occluded group.ConclusionsThis study provides preliminary evidence that physiological occlusal force may attenuate osteoclastogenesis during the early stage of tooth replantation, thereby reducing the occurrence of RRR and promoting periodontal healing.

Is genotoxicity a suitable biomarker for monitoring anabolic-androgenic steroids exposure in vivo? A systematic review and meta-analysis.

Steroids stand for a class of hormones (natural and synthetic) known to be helpful for a number of disorders. Despite the aforementioned beneficial effects of using these hormones, anabolic-androgenic steroids (AAS) are also widely abused in a non-therapeutic manner for muscle-building and strength-increasing properties that may lead to genotoxicity in different tissues. The present study aims to understand whether genotoxicity may be a suitable biomarker for AAS exposure in vivo in both experimental animal and human studies. All studies published in PubMed/Medline, Scopus, and Web of Science electronic databases that presented data on DNA damage caused by AAS were analyzed. A total of 15 articles were included in this study, and after thoroughly reviewing the studies, a total of 8 articles were classified as Strong, 6 were classified as Moderate, and only 1 was classified as Weak, totaling 14 studies being considered either Strong or Moderate. This classification makes it possible to consider the present findings as reliable. The meta-analysis data revealed a statistically significant difference in Wistar rat testis cells with AAS compared to control for tail length and % tail DNA (p < 0.001), so that the selected articles were considered homogeneous and the I2 of 0% indicated low heterogeneity. In summary, genotoxicity can be considered a suitable biomarker for monitoring AAS exposure as a result of DNA breakage and oxidative DNA damage.

A Review on Techniques of Blood Withdrawal in Experimental Small Animals

In preclinical research, experimental studies in small animals like pharmacokinetics/ toxicokinetics, serum biochemistry and hematology necessitate the biological matrix sampling to estimate drug level, serum enzyme, and blood parameters respectively. The biological matrix is withdrawn from experimental animals by employing the recommended blood collection technique and must be conducted in compliance with the ethical guidelines. It has demanded a minimal painful and least stressful procedures, which can be achieved using inhalant/parenteral anesthesia and/or proper restraint technique. This leads to better experimental outcome. The study personnel conducting blood withdrawal must be well-versed. All these procedures must be approved by IAEC protocol and should include scientist and study personnel names who are involved in animal handling and blood withdrawal. In this review, the different techniques of blood withdrawal from small animals, including collection guidelines, limit volumes and recovery periods, and advantages and disadvantages of sites used for blood collection are demonstrated and discussed.

Ethical Principles and Rules in Experimental Animal Studies: A Comprehensive Review

Ethical Principles and Rules in Experimental Animal Studies: A Comprehensive Review

Anesthesia and Analgesia Recommended for Painful Procedures in Small Animals

The non-invasive but painful and invasive procedures are frequently performed in experimental animal studies, such as blood withdrawal, route of administration, and surgical procedures. Most of these procedures commonly conducted under anesthesia and/or analgesia and should be complied with the ethical requirements. However, pain is difficult to assess in animals, therefore, it is advisable to provide the analgesics pre-emptively or if painful condition is expected. Preanesthetic like tranquilizer is generally used to reduce the stress in animal and further it also aids in the smooth induction of anesthesia and recovery. In view of this, the objective is to demonstrate and discuss the guidelines of anesthesia and analgesia in small animals, especially mouse and rat with principal focus to ethical purview. These guidelines can be applicable to the animal house facility, pharmacology laboratory, and to a large extent for veterinary care.

Feasibility of Using Serum, Plasma, and Platelet 5-hydroxytryptamine as Peripheral Biomarker for the Depression Diagnosis and Response Evaluation to Antidepressants: Animal Experimental Study.

Whether peripheral blood 5-hydroxytrptamine (5-HT) levels serve as biomarker for depression diagnosis/response evaluation has not been well determined. This work was explored to address this inconclusive issue. Animals were randomized into normal control group (NC, n = 10) and chronic unpredictable mild stress model group (CUMS-model, n = 20), respectively. Animals in CUMS-model group were subjected to chronic stress, then they were randomly subdivided into CUMS subgroup and CUMS + fluoxetine subgroup (CUMS + FLX). After FLX treatment, blood and tissues were collected. 5-HT and relevant protein expression were measured. In mice model, there was a significant increase in serum and a significant reduction in plasma 5-HT levels in CUMS-model group versus NC group, while platelet 5-HT levels change little. After FLX treatment, serum and platelet 5-HT levels were significantly decreased in CUMS + FLX subgroup, while plasma 5-HT levels had not much change versus CUMS subgroup. Chronic stress enhanced colon and platelet serotonin transporter (SERT) expression and FLX treatment mitigated SERT expression. In rats' model, there was a significant increase in serum 5-HT levels while plasma and platelet 5-HT levels showed little change in CUMS group versus NC group. After FLX treatment, serum, plasma and platelet 5-HT levels were significantly decreased in CUMS + FLX subgroup versus CUMS subgroup. The profile of relevant proteins expression changed by FLX were like those in mice. Serum 5-HT levels might serve as a potential biomarker for depression diagnosis, meanwhile serum and platelet 5-HT levels might respond to antidepressant treatment.

Feasibility and safety of dual-console telesurgery with the KangDuo Surgical Robot-1500 System using fifth-generation and wired networks: an animal experiment and sea-spanning clinical study.

To evaluate the feasibility and safety of dual-console telesurgery with the new KangDuo system in an animal experiment and clinical study. Six canine models were performed radical prostatectomy with dual-console KanDuo surgical robot-1500 (KD-SR-1500-RARP). The perioperative outcomes, physical and mental workload of the surgeon were collected. Physical workload was evaluated with surface electromyography. Mental workload was evaluated with NASA-TLX. After conducting animal experiments to verify safety of dual-console KD-SR-1500-RARP, we conducted the clinical trial using 5G and wired networks. In the animal experiment, all surgeries were performed successfully. The operative time was 80.2±32.1 min. The docking time was 2.4±0.5 min. The console time was 49.7±25.3 min. There were no perioperative complications or equipment related adverse events. All dogs can micturate after catheter removal at one week postoperatively. The mental workload was at a low level (a scale ranging from 0 to 60), which scored 15.7±6.9. Among the eight recorded muscles, the fatigue degree of the right radial flexor and left biceps was the highest two (iEMG, resection, 299.8±344 uV, 109.9±16.9 uV; suture, 849.4±1252.5 uV, 423.1±621.3 uV, respectively). In the clinical study, the console time was 136 min. The mean latency time was ≤200 ms. The data pocket loss was <1%. The operation was successfully completed without malfunctions occurring throughout the entire process. Dual-console telesurgery with the KD-SR-1500 system was shown to be feasible and safe in radical prostatectomy using 5G and wired networks.

Antinociceptive Evaluation of Antidepressants (Cita- lopram, Duloxetine and Amitriptyline) in a Mouse Model of Acute Pain

Introduction: Acute pain is a pervasive and debilitating symptom affecting the general population as well as depressed patients who suffer even more from pain and fatigue. Aims and Objectives: To evaluate the antinociceptive effect induced by antidepressants (Citalopram, Duloxetine and Amitriptyline) in acute pain induced in mice. Place and Duration of study: Animal Research Laboratory of University of Lahore. Study Period: 2018- 2019 Material and Methods: An experimental animal study was carried out on 25 albino mice. The mice were divided into 5 groups; Group 1: Positive Control (PC), Group 2: Ibuprofen (IBO) (Standard) 80mg/kg, Group3: Citalopram (CTP)3mg/kg, Group 4: Duloxetine (DXT) 25mg/kg, Group 5: Amitriptyline (AMT) 15mg/kg. Groups 2-5 were administered drugs orally through a feeding tube as per group designation while Group 1: Control was given an equivalent amount of normal saline. One hour post treatment with test drugs, all Groups (1-5) were injected 10 ml/ kg of 2% acetic acid I/P and number of writhing movements consisting of contraction of abdominal muscles leading to extension of hind limb and periodic arching of body as well as paw licking behavior were counted for 20 minutes at 5- minute interval, using hand tally counter. Degree of analgesia was calculated by using formula of percentage effectiveness. Data was entered and analyzed utilizing SPSS version 21, p?0.05 was taken as significant. Results: Ibuprofen completely eliminated the nociceptive outcome of diluted acetic acid in writhing and licking of experimental animals. Citalopram, showed maximum decreasing effect (95%-96%) in writhing in 5-20 min. However, its effect in paw licking was low. Duloxetine showed 70% to 90% decrease in writhing at 5-20 min whereas 100 % inhibition of paw-licking was observed at 5 to 20 minutes. The writhing effect of amitriptyline was 77%-90% at 5-20 min and paw licking effect was 80-90 % at 5-20 min. Conclusion: Amitriptyline, duloxetine and citalopram have significant effect in reducing the acute nociceptive pain and may be utilized as analgesic in acute state of pain.

Effects of Ficus capensis Leaves Aqueous Extracts (Lactogenic Medicinal Plant) on NMRI Mice Milk Secretion

Introduction: Ficus capensis is a plant used in traditional medicine to stimulate lactation in women and animals in Africa. However, the effects of their extracts on the mammary gland are poorly documented. The objective of the study was to evaluate the effects of Ficus capensis leaves aqueous extracts on NMRI mice milk secretion. Methodology: This was an experimental animal study using virgin female NMRI mice aged eight to ten weeks. The mice were grouped into for groups of eight mice each. Each group received one of the following products: distilled water, Galactogil™, sulpiride, aqueous extracts (AE) of capensis leaves. Data were analysed and processed using Microsoft Excel 2016 and Stata MP 16 with P ≤ .05 as the significance threshold. Results: Arborescence of the galactophorous ducts was more developed in the sulpiride lot. Galactogil™, and capensis leaves extracts treated groups showed almost equivalent arborescence with a higher tendency than the distilled water. With histological haematin- eosin staining, the ratio of galactophorous ducts containing secretions to total ducts was higher in the groups of Ficus capensis AE treated group than distilled water (P = .0001). Galactogil™, sulpiride and the group of Ficus capensis extracts each had higher levels of beta-casein in mammary tissue and average prolactinemia than distilled water (P &lt; .01). Mammary tissue stained by immunohistochemistry with anti-prolactin receptor antibodies showed more intensely labelled mammary glands in the sulpiride and Ficus capensis extracts groups. There was no statistically significant difference between average progesteronemia among the different groups. Conclusion: F. capensis leaves AE administered to virgin female NMRI mice showed lactogenic and mammogenic effects. The extracts were able to increase the nutritional quality of the milk produced, as evidenced by the increase in protein secretion.

Further developments of pubourethral ligament surgery for stress urinary incontinence.

Midurethral slings (MUS) have revolutionized the treatment of stress urinary incontinence (SUI). MUS operations work by creating a collagenous pubourethral ligament (PUL). Since 1996, more than 10 million operations have been performed worldwide. Early complications with the MUS included bowel perforations, massive retropubic hemorrhages, nerve injuries, even death. Though the invention of the transobturator tape (TOT) operation, and later, minislings, has eliminated many such complications, the most frequent complication, post-operative urinary retention, remains. MUS operations are unavailable in many countries because of expense. Low-cost surgical options discussed include the tension-free artisan minisling which uses a 10 cm × 1 cm tape inserted as a "U" below the urethra; 91% cure was achieved at 5.7 years in a study of 90 women, though it was accompanied by a 4.2% erosion rate. The more recent urethral ligament plication (ULP) is based on transperineal ultrasound studies which showed that the main cause of the SUI was elongation of weak PULs. This allowed the posterior pelvic muscles to open out the posterior urethral wall to cause SUI. Basic science collagen studies indicated that suturing PULs with No. 2 wide-bore polyester sutures would provide sufficient collagen to repair weakened PULs. Cure of SUI, when it occurred, was immediate. Reference to the original experimental animal studies indicated that collagen 1, the key structural support of PUL, had formed by 3 months. This is an optimistic sign for longer term cure, substantiated by very little deterioration after 3 months over a 12-month period in the first surgical trial (unpublished data). In conclusion, the ULP operation can be performed under local anesthesia/sedation. If it fulfils its promise for longer-term cure, SUI cure will be available for hundreds of millions of women in underdeveloped countries for a few dollars per case.

Correlations between heart sound components and hemodynamic variables

Although the esophageal stethoscope is used for continuous auscultation during general anesthesia, few studies have investigated phonocardiographic data as a continuous hemodynamic index. In this study, we aimed to induce hemodynamic variations and clarify the relationship between the heart sounds and hemodynamic variables through an experimental animal study. Changes in the cardiac contractility and vascular resistance were induced in anesthetized pigs by administering dobutamine, esmolol, phenylephrine, and nicardipine. In addition, a decrease in cardiac output was induced by restricting the venous return by clamping the inferior vena cava (IVC). The relationship between the hemodynamic changes and changes in the heart sound indices was analyzed. Experimental data from eight pigs were analyzed. The mean values of the correlation coefficients of changes in S1 amplitude (ΔS1amp) with systolic blood pressure (ΔSBP), pulse pressure (ΔPP), and ΔdP/dt during dobutamine administration were 0.94, 0.96, and 0.96, respectively. The mean values of the correlation coefficients of ΔS1amp with ΔSBP, ΔPP, and ΔdP/dt during esmolol administration were 0.80, 0.82, and 0.86, respectively. The hemodynamic changes caused by the administration of phenylephrine and nicardipine did not correlate significantly with changes in the heart rate. The S1 amplitude of the heart sound was significantly correlated with the hemodynamic changes caused by the changes in cardiac contractility but not with the variations in the vascular resistance. Heart sounds can potentially provide a non-invasive monitoring method to differentiate the cause of hemodynamic variations.

Effects of Saikosaponin-A on Insulin Resistance in Obesity: Computational and Animal Experimental Study.

Obesity is known to be associated with increased inflammation and dysregulated autophagy, both of which contribute to insulin resistance. Saikosaponin-A (SSA) has been reported to exhibit anti-inflammatory and lipid-lowering properties. In this research, we employed a combination of computational modeling and animal experiments to explore the effects of SSA. Male C57BL/6 mice were categorized into four groups: normal diet, high-fat diet (HFD), HFD + atorvastatin 10 mg/kg, and HFD + SSA 10 mg/kg. We conducted oral glucose and fat tolerance tests to assess metabolic parameters and histological changes. Furthermore, we evaluated the population of Kupffer cells (KCs) and examined gene expressions related to inflammation and autophagy. Computational analysis revealed that SSA displayed high binding affinity to tumor necrosis factor (TNF)-α, nuclear factor (NF)-κB, fibroblast growth factor 21 (FGF21), and autophagy-related 7 (ATG7). Animal study demonstrated that SSA administration improved fasting and postprandial glucose levels, homeostatic model assessment of insulin resistance (HOMA-IR) index, as well as triglyceride, free fatty acid, total cholesterol, low-density lipoprotein cholesterol (LDL-C)-cholesterol, and high-density lipoprotein cholesterol (HDL-C)-cholesterol levels in HFD-fed mice. Moreover, SSA significantly reduced liver weight and fat accumulation, while inhibiting the infiltration and M1 activation of KCs. At the mRNA level, SSA downregulated TNF-α and NF-κB expression, while upregulating FGF21 and ATG7 expression. In conclusion, our study suggests that SSA may serve as a therapeutic agent for addressing the metabolic complications associated with obesity. This potential therapeutic effect is attributed to the suppression of inflammatory cytokines and the upregulation of FGF21 and ATG7.

General Principles, Designs, and Statistical Analyses in Experimental Animal Studies

Research using animals contributes significantly to many research and development studies, especially in the biomedical field. Within the scope of the study, conducting animal experiments in accordance with scientific principles and ensuring the ethical use and welfare of animals are issues that should be taken into consideration. In this context, the scientific contribution to be achieved by conducting these studies in line with scientific and ethical principles will be directly proportional. In many studies, while investigating the biological significance, it is seen that the effects of many factors are ignored, the answer to the biological question is investigated with simple experimental designs, or the accurate statistical analyses are not chosen. Therefore, in this study, the principles that a researcher planning an animal study should follow within the scope of the research (animal ethics, 3R, and other R rules, determination of sample size, randomization, and blinding) are briefly mentioned. Then, completely randomized design, regression design, split-unit design, hierarchical (nested) design, mixed effects design, and appropriate statistical analyses for these designs, which are thought to be useful in these studies, are discussed. It is thought that this review will be useful as it contains important summative information that will guide all researchers in planning animal studies accurately and quickly.

Factor XI inhibition in patients with acute coronary syndrome.

A hypercoagulable condition is typical of patients with acute coronary syndrome and is a determining factor in the genesis of recurrent ischaemic events. Modern pharmacological therapies consisting of antiplatelets and anticoagulants derive their rationale for use on the pathophysiological mechanisms most commonly associated with myocardial infarction (MI); they have contributed to reducing the ischaemic risk of these patients, but left ample room for improvement. In particular, trials that have studied the association of an anticoagulant with antiplatelet drugs have provided promising results in terms of efficacy, but highlighted a significant bleeding risk. Evidence derived from experimental animal and epidemiological studies has shown how factor XI (FXI) deficiency is associated with a reduction in thrombotic events but with modest bleeding. These data added to the role that FXI plays in the coagulation cascade constituted an incipit for the pharmacological attempt to decouple thrombosis from haemostasis by means of the inhibition of this factor. The theoretical assumption that FXI inhibitor drugs may be able to reduce the ischaemic risk without significantly increasing the haemorrhagic risk makes these compounds a potential therapeutic aid for patients in secondary prevention after acute MI. To date, on these patients, we only have data from a Phase 2 trial, PACIFIC-AMI (Study to Gather Information About the Proper Dosing and Safety of the Oral FXIa Inhibitor BAY 2 433 334 in Patients Following an Acute Heart Attack). In this study, the primary endpoint-represented by the Bleeding Academic Research Consortium (BARC) composite of Type 2, 3, or 5 bleeding-showed no significant differences between the various doses of asundexian tested (10, 20, and 50 mg quoque die), and between these and placebo (asundexian all doses vs. placebo: hazard ratio, 0.98; 90% confidence interval, 0.71-1.35). The data on efficacy, however, showed neutral results, but it should be noted that the study did not have the adequate statistical power to evaluate this outcome. Valuable information could, therefore, derive in the future from the ongoing Phase 3 trial with milvexian, LIBREXIA-ACS (A Study of Milvexian in Participants After a Recent Acute Coronary Syndrome) and from any future studies that could be started by testing different molecules.

PROTECTIVE EFFECTS OF SILYMARIN AND CHOLCALCIFEROL ON ISONIAZID INDUCED HEPATOTOXICITY IN MICE

Background: Hepatotoxicity is the gravest concern associated with use of anti-tuberculous drugs. The objective of this study was to evaluate the individual and combined hepatoprotective effect of silymarin and cholecalciferol in isoniazid induced hepatotoxicity. Methods: This animal experimental study was conducted at the Department of Pharmacology &amp; Therapeutics, and Multidisciplinary Research Laboratory, Islamic International Medical College, in collaboration with National Institute of Health, Islamabad, Pakistan. Fifty adult Balb-C mice were included in this study. They were distributed into 5 groups. Each group contained 10 mice. Group 1 was normal control, Group 2 disease control, and Group 3, 4, and 5 were experimental groups. Except Group 1, all other groups were given isoniazid (150 mg/Kg) and only Group 2 was not fed with any drugs. Group 3 received silymarin (50 mg/Kg dissolved in physiological saline) through intragastric gavage for 28 days. Group 4 was given Vitamin D (1,000 IU/Kg) for 28 days. Group 5 was given isoniazid (150 mg/Kg) along with silymarin and Vitamin D for 28 days. Serum ALT and bilirubin levels were estimated on day 0, 14, and 28. Results: As compared to Group 2, Group 3 to Group 5 showed a lower rise in serum ALT and bilirubin (p&lt;0.001). Group 4 and 5 showed significantly reduced biochemical markers (ALT and bilirubin) (p=0.001). Conclusion: Silymarin and cholecalciferol effectively and synergistically ameliorate hepatotoxicity induced by isoniazid. Silymarin offers better hepatoprotection than cholecalciferol in isoniazid induced hepatotoxicity. Pak J Physiol 2024;20(1):37-40

POTENTIAL RESTORATION OF FATTY LIVER ENZYMES WITH STEVIA REBAUDIANA

Background: Obesity is a prevalent issue affecting a growing number of individuals globally. It is a systemic disorder with complications and co-morbidities. Non-alcoholic fatty liver disease (NAFLD) is a disastrous outcome of metabolic syndrome induced by obesity. Objective of this study was to find the effect of Stevia leaves in restoration of liver function tests (ALT, AST, ALP, and Bilirubin) in obese Sprague Dawley rats. Methods: This animal experimental study was carried out in the Physiology Department, in collaboration with the Pathology Department of Islamabad Medical and Dental College, and National Institute of Health, Islamabad on 90 healthy male Sprague Dawley rats over a period of 14 weeks. The animals were divided randomly into three groups of 30 rats each. Group 1 was given normal diet while Groups 2 and Group 3 were given high fat diet. Stevia leaves were further added for six weeks in the diet of Group 3. Results: High fat diet induced NAFLD in rats was ameliorated significantly on treatment with stevia. On comparison of liver function tests in obese control with stevia treated group, the values of ALT and AST were significantly decreased (p&lt;0.05). However, ALP and bilirubin were not decreased significantly. Conclusion: Stevia rebaudiana exerts hepatoprotective effect in restoring liver damage due to high fat diet induced NAFLD in Sprague Dawley rats. Pak J Physiol 2024;20(1):22-5