Journal of pharmacological and toxicological methods最新文献

{"title":"Molecular imaging of excitability difference between alkaloids/salts (nicotine, nicotinic benzoate, caffeine and arecoline hydrobromide)","authors":"Dawei Yan ,&nbsp;Xiaomin Liu ,&nbsp;Yihan Gao ,&nbsp;Xiaonan Li ,&nbsp;Xiabin Chen ,&nbsp;Yiting Qian ,&nbsp;Saijing Zheng ,&nbsp;Yi Shen","doi":"10.1016/j.vascn.2024.107569","DOIUrl":"10.1016/j.vascn.2024.107569","url":null,"abstract":"<div><div>Comparison of the excitability of four different alkaloids/salts, including nicotine, nicotinic benzoate, caffeine and arecoline hydrobromide. Based on positron emission tomography (PET) imaging and ¹⁸F-Fallypride, a novel technique for measuring alkaloid/salt excitability in SD rats was developed. Different doses and types of alkaloids/salts were administered to the SD rats in a single nebulised inhalation.</div><div>The results showed that: (1) PET imaging technology can detect the excitability intensity of SD rats after single inhalation of alkaloids/salts in non-invasive real time and the optimal PET scanning time of four different alkaloids/salts (nicotine, nicotinic benzoate, caffeine and arecoline hydrobromide) were slightly different. (2) The excitatory saturation effect of four alkaloids/salts was observed in SD rats after single inhalation and the saturation effect doses of nicotine, nicotine benzoate, caffeine and arecine hydrobromide were 0.063 mg/kg, 0.075 mg/kg, 0.33 mg/kg and 0.075 mg/kg, respectively. (3) In the case of single inhalation of the same dose of four alkaloids/salts, male SD rats inhaled arecoline hydrobromide with the strongest excitability, while female SD rats inhaled nicotinic benzoate. A PET method for noninvasive real-time detection of alkaloid/salt excitability in SD rats was established.</div><div>The finding of an excitatory saturation effect for four alkaloids/salts (nicotine, nicotinic benzoate, caffeine and arecoline hydrobromide) and the presence of excitatory intensity and gender differences at the same dose of inhalation of four alkaloids/salts, which provide a new theoretical basis for determiningthe content of alkaloids/salts.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"130 ","pages":"Article 107569"},"PeriodicalIF":1.3,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142515417","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In-situ polyherbal gel as biomedicine in the management of Alzheimer's disease: Understanding ameliorative potential in Trimethyltin induced neurodegeneration","authors":"Sneha Latha Diddi ,&nbsp;Sathiyanarayanan Lohidasan ,&nbsp;Arulmozhi S ,&nbsp;Vividha Dhapte-Pawar ,&nbsp;Kakasaheb R. Mahadik","doi":"10.1016/j.vascn.2024.107567","DOIUrl":"10.1016/j.vascn.2024.107567","url":null,"abstract":"<div><div>Alzheimer's disease (AD), classified as neurodegenerative disorder that progresses over a period of time, is characterized by intracellular neurofibrillary tangles and extracellular amyloid plaques. This present research work was designed to develop a polyherbal gel for the treatment of AD. This research study is aimed to confirm the impact and validation of polyherbal gel on tauopathy and neurodegeneration that had been induced by intraperitoneal trimethyltin (TMT) injection to rats.</div><div>Polyherbal loaded gel was prepared by cold method, and characterized for gel strength, viscosity, permeation and pH. Subsequently, 5 marker based standardized plant materials of <em>Kalyanka ghrita</em> were incorporated in gellan gum and xanthan gum. Finally, an <em>in-vivo</em> investigation employing rats with TMT-induced neurological disease were used to assess the efficacy of the optimized gel.</div><div>On day 7, the Wistar rats received intraperitoneal injections of TMT. From day 14 to day 35, the corresponding groups received intranasal administration of polyherbal gel. In addition to the molecular parameters such as brain acetyl cholinesterase activity, BDNF (Rat brain derived neurotropic factor), protein phosphatase 2 A, antioxidant parameters, and oxidative stress markers, the behavioral parameters were also determined. Studies were conducted on the brain's monoamine levels and histology.</div></div><div><h3>Results</h3><div>Higher permeation over the nasal mucosa was demonstrated by the optimized <em>In-situ</em> polyherbal gel. Significant improvement in cognition was observed from the reduced escape latency, longer paths, and increased social or novel object recognition tests post polyherbal gel treatment. A documented HPLC technique helped in optimization and standardization of the polyherbal gel. The polyherbal treatment groups exhibited a considerable rise in the levels of monoamines, including norepinephrine, dopamine, and 5-hydroxy tryptamine.</div></div><div><h3>Conclusion</h3><div>According to the current study, treating Alzheimer's disease (AD) with a polyherbal gel formulation may be a viable option for successful therapy.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"130 ","pages":"Article 107567"},"PeriodicalIF":1.3,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142407326","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of 4 quantification methods for monitoring 16 antidepressant drugs and their metabolites in human plasma by LC-MS/MS","authors":"Lingjiao Wang ,&nbsp;Mengqiang Zhao ,&nbsp;Jiao Li ,&nbsp;Hongtao Xu ,&nbsp;Yang Song ,&nbsp;Yuanyuan Zhao ,&nbsp;Jing Yu ,&nbsp;Chunhua Zhou","doi":"10.1016/j.vascn.2024.107568","DOIUrl":"10.1016/j.vascn.2024.107568","url":null,"abstract":"<div><h3>Background</h3><div>Therapeutic drug monitoring for antidepressants (ADs) is vital due to the potentially serious consequences and disputes related to medical events. Therefore, we created a quick and convenient analysis way for separation and quantification of ADs.</div></div><div><h3>Methods</h3><div>To ensure quantitative stability, we divided the 16 ADs or their metabolites into 4 pools (AD1-AD4), considering the hospital frequency that the clinician prescribed, the physicochemical properties of medicines, and the calibration range of selected ADs. After precipitation with methanol, the analytes were eluted for at least 3.5 min on a BEH C₁₈ analytical column by different gradient elution methods. <strong>Results:</strong> The LLOQ and LOD were 1.25–10 ng/mL and 0.42–5 ng/mL, respectively. High precision (&lt;12 %) and accuracy (87.07–111.47 %) were demonstrated by quality control samples both within and between days. All the compounds were stable at room temperature and within −80 °C.</div></div><div><h3>Conclusion</h3><div>The method is of wide clinical and laboratory interest due to simpler sample cleanup, shorter chromatographic run times, and wider calibration range compared to other methods.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"130 ","pages":"Article 107568"},"PeriodicalIF":1.3,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142396419","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative toxicity assessment of selected nanoparticles using different experimental model organisms","authors":"Srishti Parashar ,&nbsp;Sheetal Raj ,&nbsp;Priyanka Srivastava ,&nbsp;Abhishek Kumar Singh","doi":"10.1016/j.vascn.2024.107563","DOIUrl":"10.1016/j.vascn.2024.107563","url":null,"abstract":"<div><div>Nanoparticles are microscopic particles ranging in size from one to one hundred nanometers. Due to their extensive features, nanoparticles find widespread use in various fields worldwide, including cosmetics, medical diagnosis, pharmaceuticals, food products, drug delivery, electronic devices, artificial implants, and skincare. However, their unique characteristics have led to high demand and large-scale manufacturing, resulting in adverse impacts on the environment and bioaccumulation. Researchers have been exploring issues related to the environmental toxicity resulting from the high production of selected nanoparticles. This review discusses and addresses the adverse impacts of highly produced nanoparticles such as Carbon Nanotubes, Silica, Titanium dioxide, Zinc Oxide, Copper oxide, and Silver nanoparticles on different in vivo<em>,</em> in vitro, alternate invertebrate models, and plant models. Summarizing in vivo research on rats, rabbits, and earthworms, the review reveals that nanoparticles induce cytotoxicity, embryotoxicity, and DNA damage, primarily targeting organs like the brain, liver, kidney, and lungs, leading to nephron, neuro, and hepatotoxicity. Studying the effects on alternative models like zebrafish, <em>Caenorhabditis elegans</em>, <em>Drosophila</em>, sea urchins, and <em>Saccharomyces cerevisiae</em> demonstrates genotoxicity, apoptosis, and cell damage, affecting reproduction, locomotion, and behavior. Additionally, research on various cell lines such as HepG2, BALB/c 3 T3, and NCL-H292 during in vitro studies reveals apoptosis, increased production of reactive oxygen species (ROS), halted cell growth, and reduced cell metabolism. The review highlights the potentially adverse impacts of nanoparticles on the environment and living organisms if not used sustainably and with caution. The widespread use of nanoparticles poses hazards to both the environment and human health, necessitating appropriate actions and measures for their beneficial use. Therefore, this review focuses on widely used nanoparticles like zinc, titanium, copper, silica, carbon nanotubes, and silver, chosen due to their environmental toxicity when excessively used. Environmental toxicity of air, water, and soil is evaluated using environmentally relevant alternative animal models such as Drosophila, zebrafish, earthworms, etc., alongside in vivo and in vitro models, as depicted in the graphical abstract.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"130 ","pages":"Article 107563"},"PeriodicalIF":1.3,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142368062","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluating the proarrhythmic risk of delayed-action compounds in serum free cell culture conditions; serum-starvation accelerates/amplifies the effect of probucol on the KCNQ1 + KCNE1 channel","authors":"Kenny M. Van Theemsche ,&nbsp;Lisse Frans ,&nbsp;Dieter V. Van de Sande ,&nbsp;Evelyn Martinez-Morales ,&nbsp;Dirk J. Snyders ,&nbsp;Alain J. Labro","doi":"10.1016/j.vascn.2024.107566","DOIUrl":"10.1016/j.vascn.2024.107566","url":null,"abstract":"<div><div>In vitro testing procedures for evaluating acute effects of compound on ion channels, utilizing heterologous expression systems (HES), are well-established, while slowly manifesting delayed effects remain challenging to detect. For this, immortalized HES are exposed to the compounds for a longer time, in general 24 h. As these cells proliferate every 12–20 h, we evaluated if the proliferation status, and by extension cell metabolism, influences the delayed compound response. The intervention of halting cell proliferation by excluding serum from the culturing medium was evaluated on CHO cells, stably expressing the KCNQ1 + KCNE1 channel complex that mediates the slow delayed rectifier potassium current (I_ks). No abnormal changes in KCNQ1 + KCNE1 current were observed upon serum-starvation, except for a negative shift in the voltage dependence of channel activation (GV-curve) after 72 h. The delayed effect of probucol, a compound reported to interfere with I_ks expression, was evaluated after 24 and 72 h of incubation. In serum-free conditions the inhibitory effect of probucol was increased fourfold after 24 h, compared to serum supplemented conditions. After 72 h, the current inhibition was similar between both culture conditions. Besides decreasing current expression, probucol shifted the GV-curve more positive combined with a shallower voltage response, changes that were more pronounced in serum-depleted conditions. The results indicated that serum-starvation had no substantial effect on the KCNQ1 + KCNE1 current in the tested CHO cells, but it amplified or accelerated the response to probucol, suggesting that halting cell proliferation is a method for enhancing the detection of delayed compound effects in HES.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"130 ","pages":"Article 107566"},"PeriodicalIF":1.3,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142368063","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhibition of hERG K channels by verapamil at physiological temperature: Implications for the CiPA initiative","authors":"Ashley A. Johnson,&nbsp;Matthew C. Trudeau","doi":"10.1016/j.vascn.2024.107562","DOIUrl":"10.1016/j.vascn.2024.107562","url":null,"abstract":"<div><div>The Comprehensive in vitro Proarrhythmia Assay (CiPA) initiative reassesses using the inhibition of hERG potassium channels by drugs as the major determinant for the potential to cause drug-induced Torsades de Pointes (TdP) cardiac arrhythmias. Here we report our findings on the next phase of CiPA: Determination of hERG inhibitory properties using the standard CiPA-defined data acquisition protocol, here called the standard protocol, at physiological temperature (37 degrees Celsius). To do this, we measured inhibition of hERG1a potassium channels stably expressed in HEK293 cells by the small molecule verapamil, using manual whole-cell patch-clamp electrophysiology recordings with the standard protocol, which is characterized, in part, by a series of 10 s duration voltage steps to 0 mV, ultimately leading to a cumulative recording time of approximately 30 min. Using the standard protocol, we measured an IC₅₀ for verapamil of 225 nM, a Hill coefficient of 1, and time constant of inhibition at 0 mV of 0.64 s. But, using the standard protocol resulted in a very low (5 %) experimental success rate per cell, which had low practicality for future experiments. To address the 5 % success rate, we generated a revised protocol characterized, in part, by a series of 3 s duration voltage steps to 0 mV, leading to a cumulative recording time of approximately 10 min. Using the revised protocol, we found an IC₅₀ for verapamil of 252 nM, a Hill coefficient of 0.8, and time constant of inhibition at 0 mV of 0.67 s. The values measured with the revised protocol were similar to those measured using the standard protocol and, furthermore, our success rate using the revised protocol rose to 25 %, an increase of 5-fold over the standard protocol, and more in line with the success rate for biophysical studies. In summary, we captured key pharmacological data for subsequent analysis in CiPA using a revised protocol with an increased success rate and an overall enhanced feasibility and practicality. We propose that the revised protocol may be more pragmatic for generation of some hERG channel drug inhibition data for CiPA and other regulatory sciences.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"130 ","pages":"Article 107562"},"PeriodicalIF":1.3,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142335349","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Overcoming obstacles in three-dimensional cell culture model establishment: Approaches for growing A549 non-small cell lung cancer spheroids using a clinostat system","authors":"Charity M. Mabela ,&nbsp;Chrisna Gouws ,&nbsp;Wihan Pheiffer","doi":"10.1016/j.vascn.2024.107564","DOIUrl":"10.1016/j.vascn.2024.107564","url":null,"abstract":"<div><h3>Introduction</h3><div>Non-small cell lung cancer (NSCLC) accounts for 80–85 % of lung cancer cases globally. And the A549 cell line is widely used in pharmacological and toxicity screening. Due to its popularity as a NSCLC model, it was inevitable that three-dimensional (3D) cultures of A549 cells would be established. 3D models increase physiological relevance, and their advanced structure allows researchers to obtain more translatable and reliable results. However, establishing this cell line as a 3D model may come with challenges, like clumping.</div></div><div><h3>Methods</h3><div>In this study, A549 spheroids were established using a clinostat-based rotating bioreactor system and were characterised in terms of morphology, planimetry, and viability.</div></div><div><h3>Results</h3><div>The main challenge faced included continuous aggregation of the spheroids, which constrained growth and development. This challenge was successfully overcome by supplementation with ascorbic acid, foetal bovine serum coating, and minimising handling, and a NSCLC mini-tumour model was established and semi-characterised. The spheroids survived for 25 days and had a significant increase in growth.</div></div><div><h3>Conclusion</h3><div>The A549 spheroid model cultured in a clinostat-based microgravity system was shown to be stable, viable, and suitable to be used in pharmacological and toxicological investigations.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"130 ","pages":"Article 107564"},"PeriodicalIF":1.3,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142335348","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A general and rapid LC-MS/MS method for simultaneous determination of voriconazole, posaconazole, fluconazole, itraconazole and hydroxyitraconazole in IFI patients","authors":"Wenwen Xia ,&nbsp;Shun Chen ,&nbsp;Yunlei Yun ,&nbsp;Lili Cui ,&nbsp;Zhipeng Wang ,&nbsp;Juanjuan Hou ,&nbsp;Mao Tang ,&nbsp;Chen Bu ,&nbsp;Shouhong Gao ,&nbsp;Rongzi Shao ,&nbsp;Xia Tao","doi":"10.1016/j.vascn.2024.107565","DOIUrl":"10.1016/j.vascn.2024.107565","url":null,"abstract":"<div><h3>Objective</h3><div>To establish a rapid and universal quantitative liquid chromatography tandem mass spectrometry (LC-MS/MS) method for measuring the exposure levels of five triazole antifungal drugs in human plasma, including voriconazole, fluconazole, posaconazole, itraconazole, and hydroxyitraconazole.</div></div><div><h3>Methods</h3><div>A triple quadrupole mass spectrometer operating in positive ionization mode was used to detect the analyte, and multiple reaction monitoring mode was employed to gather data. The mobile phase included 0.05 % formic acid in water (phase A) and acetonitrile (phase B). The analytes were separated on an Agilent EclipsePlusC₁₈ RRHD column (30 × 50 mm, 1.8 μm) using gradient elution. The flow rate was 0.3 mL/min with the column temperature set at 35 °C. The acetonitrile was used to pretreat the plasma sample, and the itraconazole-D5 and hydroxyitraconazole-D5 were utilized as the internal standards.</div></div><div><h3>Results</h3><div>The calibration range was from 100 to 10,000 ng/mL for posaconazole, itraconazole, and hydroxyitraconazole, from 200 to 20,000 ng/mL for fluconazole and from 50 to 5000 ng/mL for voriconazole, with linear correlation coefficients more than 0.99 for all regression curves. The intra- and inter-day accuracy and precision of the method were within ±15 %. The mean extraction recovery of all the analytes ranged from 74.32 % to 117.83 %, and the matrix effect was from 72.54 % to 111.2 %. The results of stability fell into the scope of ±15 % deviation.</div></div><div><h3>Conclusion</h3><div>This newly developed method is sensitive, simple, and robust, and successfully applied in determining triazole antifungal drugs in plasma from 66 IFI patients to provide reference for safe and effective drug administration in clinical practice.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"130 ","pages":"Article 107565"},"PeriodicalIF":1.3,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142335346","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assessment of antiemetic activity of dihydrocoumarin: In vivo and in silico approaches on receptor binding affinity and modulatory effects","authors":"Md. Abu Saim ,&nbsp;Md. Shimul Bhuia ,&nbsp;Tanzila Akter Eity ,&nbsp;Raihan Chowdhury ,&nbsp;Nowreen Tabassum Ahammed ,&nbsp;Siddique Akber Ansari ,&nbsp;Kazi Nadim Hossain ,&nbsp;Afroza Akter Luna ,&nbsp;Md. Hanif Munshi ,&nbsp;Muhammad Torequl Islam","doi":"10.1016/j.vascn.2024.107561","DOIUrl":"10.1016/j.vascn.2024.107561","url":null,"abstract":"<div><div>Dihydrocoumarin (DCN) is a natural compound widely used in the flavor industry and has antioxidant and anti-inflammatory properties. However, its potential antiemetic effects on gastrointestinal disturbances remain untested. This study emphasizes assessing the antiemetic properties of the natural aromatic compound DCN using copper sulfate (CuSO₄.5H₂O)-induced emetic model on chicks, and an <em>in silico</em> approach was also adopted to estimate the possible underlying mechanisms. Two doses (25 and 50 mg/kg b.w.) of DCN and several referral drugs considered positive controls (PCs), including domperidone (6 mg/kg), hyoscine (21 mg/kg), aprepitant (16 mg/kg), diphenhydramine (10 mg/kg), and ondansetron (5 mg/kg), were orally administered to chicks. The vehicle was provided as the control group. Co-treatments of DCN with referral drugs were also provided to chicks to evaluate the modulatory action of the test compound. According to the results, DCN delayed the emetic onset and decreased the frequency of retches in a dose-dependent manner compared to the vehicle group. DCN (50 mg/kg) represented a notable delayed latency period (61.17 ± 4.12 s) and a diminished number of retchings (17.67 ± 1.82 times) compared to the control group. Further, in the co-treatments, DCN increased the latency period and reduced the number of retches, except for domperidone. In the <em>in silico</em> investigation, DCN showed notable binding affinity toward the D₂ (−7 kcal/mol), H₁ (−7.5 kcal/mol), and M₅ (−7 kcal/mol) receptors in the same binding site as the referral ligand. Our research indicates that DCN has mild antiemetic properties by interacting with the D₂, H₁, and M₅ receptors. Therefore, several pre-clinical and clinical studies are necessary to assess the effectiveness and safety profile of this food ingredient.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"130 ","pages":"Article 107561"},"PeriodicalIF":1.3,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142323413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative analysis of high-throughput RNA extraction kits in Naïve Non-Human Primate (NHP) tissues for downstream applications utilizing Xeno Internal Positive Control (IPC)","authors":"Ruwini D. Rajapaksha,&nbsp;Catherine Brooks,&nbsp;Adriana Rascon,&nbsp;Adam Fadem,&nbsp;Ivy Nguyen,&nbsp;Philip J. Kuehl,&nbsp;John T. Farmer","doi":"10.1016/j.vascn.2024.107549","DOIUrl":"10.1016/j.vascn.2024.107549","url":null,"abstract":"<div><p>Ribonucleic acid (RNA) extraction and purification play pivotal roles in molecular biology and cell and gene therapy, where the quality and integrity of RNA are critical for downstream applications. Automated high-throughput systems have gained interest due to their potential for scalability and reduced labor requirements compared to manual methods. However, ensuring high-throughput capabilities, reproducibility, and reliability while maintaining RNA yield and purity remains challenging.</p><p>This study evaluated and compared the performance of four commercially available high-throughput magnetic bead-based RNA extraction kits across six types of naïve non-human primate (NHP) tissue matrices: brain, heart, kidney, liver, lung, and spleen. The assessment focused on RNA purity, yield, and extraction efficiency (EE) using Xeno Internal Positive Control (IPC) spiking.</p><p>Samples (∼50 mg) were homogenized via bead-beating and processed according to the manufacturer's protocol on the KingFisher Flex platform in eight replicates. RNA purity and yield were measured using a NanoDrop® spectrophotometer, while EE was evaluated via real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR).</p><p>The findings indicate consistent high RNA purity across all tested extraction kits, yet substantial variation in RNA yield. Extraction efficiency exhibited variations across tissue types, with decreasing trends observed from brain to lung tissues. These results underscore the importance of careful kit selection and method optimization for achieving reliable downstream applications. The MagMAX™ <em>mir</em>Vana™ Total RNA Isolation Kit stands out as the most accurate and reproducible, making it the preferred choice for applications requiring high RNA quality and consistency. Other kits, such as the Maxwell® HT simplyRNA Kit, offer a good balance between cost and performance, though with some trade-offs in precision. These findings highlight the importance of selecting the appropriate RNA isolation method based on the specific needs of the research, underscoring the critical role of accurate nucleic acid extraction in gene and cell therapy research.</p><p>In conclusion, this study highlights the critical factors influencing RNA extraction performance, emphasizing the need for researchers and practitioners to consider both kit performance and tissue characteristics when designing experimental protocols. These insights contribute to the ongoing efforts to enhance the reproducibility and reliability of RNA extraction methods in molecular biology and cell/gene therapy applications.</p></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"129 ","pages":"Article 107549"},"PeriodicalIF":1.3,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1056871924000595/pdfft?md5=710694baa0fad90506a661d8de218c74&pid=1-s2.0-S1056871924000595-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142142208","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}