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Characterization of Forced Degradation Products of Netarsudil: Optimization and Validation of a Stability-Indicating RP-HPLC Method for Simultaneous Quantification of Process-Related Impurities. 耐塔舒地尔强制降解产物的表征:同时定量耐塔舒迪尔工艺相关杂质的稳定性指示RP-HPLC方法的优化和验证
IF 1.8 Q3 PHARMACOLOGY & PHARMACY Pub Date : 2024-07-12 DOI: 10.4274/tjps.galenos.2023.99148
Venkateswara Rao Anna, Bodasingi Sai Kumar, Jammu Harish, Bhagya Kumar Tatavarti, Tamma Eswarlal

Objectives: The aim of this study is to examine resolution, identification, and characterization of forced degradation products of netarsudil by liquid chromatography-tandem mass spectrometry by validating a simple and sensitive high-performance liquid chromatography method for the resolution, identification, and quantification of two process-related impurities in netarsudil.

Materials and methods: Chromatographic separation was accomplished on a ZORBAX Eclipse XDB C18 (250 x 4.6 mm; 5 µ id) column at room temperature as the stationary phase and 257 nm as the detector wavelength with the mobile phase consisting of acetonitrile, methanol, and pH 4.6 phosphate buffer in 45:35:20 (v/v) at 1.0 mL/min flow rate in isocratic elution.

Results: The method reported very sensitive detection limits of 0.008 µg/mL for impurity 1 and 0.003 µg/mL for impurity 1. The method produces a calibration curve linear in the concentration level of 25-200 for netarsudil and 0.025-0.2 µg/mL for impurities. The proposed method gives acceptable results for other validation parameters such as accuracy, precision, ruggedness, and robustness. The drug was subjected to various stress conditions such as acid, base, peroxide, and thermal and ultraviolet light to investigate the stability-indicating ability of the method. Considerable degradation was observed in stress studies, and the degradation products were well resolved from process-related impurities. The characterization of degradation products was performed on the basis of collision-induced dissociation mass spectral data, and the possible structures of the six degradation compounds of netarsudil were proposed.

Conclusion: The outcomes of other validation studies were likewise satisfactory and proven adequate for the regular analysis of netarsudil and its process-related impurities in bulk drug and pharmaceutical dosage forms and can also be applied for the evaluation of the stress degradation mechanism of netarsudil.

摘要
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引用次数: 0
Effect of Nutrition on Drug-Induced Liver Injury: Insights from a High-Fat Diet Mouse Model. 营养对药物性肝损伤的影响:来自高脂饮食小鼠模型的见解
IF 1.8 Q3 PHARMACOLOGY & PHARMACY Pub Date : 2024-07-12 DOI: 10.4274/tjps.galenos.2023.23855
Murali Badanthadka, Vinitha D'Souza, Meghashree Shetty, Varsha Augustin, Madhura Rathnakar Jalajakshi, Mamatha Bangera Sheshappa, Vijayanarayana Kunhikatta

Objectives: Literature suggests that a high-fat diet (HFD) potentially increases the risk of chemical/drug-induced toxicity after an acute overdose. Drug/chemical-induced hepatotoxicity has been well studied, and the mechanism that regulates this toxicity has been extensively examined using different experimental animal models. Our study focuses on drug-induced hepatotoxicity in HFD-fed female Balb/C mice. This study addresses the effect of nutrition on the magnitude of acetaminophen (APAP)-induced hepatotoxicity at different time intervals.

Materials and methods: Female Balb/C mice, after the weaning period separated into two different groups, normal diet (ND) and HFD receiving groups; after 15 weeks, they were dosed with a single dose (300 mg/kg per os (p.o.) of APAP. Blood samples were collected at different time intervals (0, 6 and 24 hours), and liver samples were collected at the end time point. Liver injury parameters [alanine aminotransferase (ALT) and aspartate aminotransferase (AST)], antioxidant assay (sodium dismutase, glutathione, and catalase), and histopathology study were conducted. Pharmacokinetic (PK) analysis was done using the RP-HPLC system and Phoenix WinNonlin 8.3 software.

Results: APAP-induced liver injury decreased AST and ALT in the HFD group compared with the ND group at 6 and 24 hours (p < 0.01 and p < 0.001), respectively. Antioxidant enzyme levels remained constant in the HFD group, whereas histopathology showed remarkable changes. The PK's of APAP in HFD indicate lower plasma concentrations of APAP (p < 0.05), with two-fold higher clearance and volume of distribution.

Conclusion: HFD significantly reduced susceptibility to APAP-mediated liver injury in Balb/C mice compared with ND mice. Our study mimics the clinical scenario where the same dose of the drug is prescribed to the normal and obese population. Our results suggest the potential need for dose titration to assess an individual's nutritional state in a clinical scenario.

目的:文献表明,高脂肪饮食(HFD)可能会增加急性服药过量后化学/药物诱导毒性的风险。药物/化学物质诱导的肝毒性已经得到了很好的研究,并且使用不同的实验动物模型对调节这种毒性的机制进行了广泛的研究。我们的研究重点是HFD喂养的雌性Balb/C小鼠的药物诱导的肝毒性。本研究探讨了营养在不同时间间隔对APAP诱导的肝毒性程度的影响。材料和方法:断奶后的雌性Balb/c小鼠分为正常饮食(ND)和高脂饮食(HFD)两组,15周后给予单剂量(300mg/kg,p.o.)的对乙酰氨基酚(APAP)。在不同的时间间隔(0、6和24小时)采集血液样本,在结束时间点采集肝脏样本。进行肝损伤参数(ALT和AST)、抗氧化剂测定(SOD、GSH和过氧化氢酶)和组织病理学研究。采用RP-HPLC法和Phoenix WinNonlin 8.3软件进行药代动力学分析。结果:与正常饮食(ND)组相比,APAP诱导的肝损伤降低了HFD组的AST、ALT。HFD组的抗氧化酶水平保持不变,而组织病理学表现出显著变化。APAP在HFD中的药代动力学表明,它具有较低的APAP血浆浓度,具有两倍高的清除率和分布体积。结论:与ND小鼠相比,HFD降低了Balb/C小鼠对APAP介导的肝损伤的易感性。我们的研究模拟了正常和肥胖人群服用相同剂量药物的临床情况。我们的研究结果表明,在临床情况下,可能需要对个体的营养状态进行剂量滴定。
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引用次数: 0
Screening of Antimicrobial, Antibiofilm, and Cytotoxic Activities of Some Medicinal Plants from Balıkesir Province, Türkiye: Potential Effects of Allium paniculatum Flower. 土耳其Balıkesir省部分药用植物的抗菌、抗菌膜和细胞毒性筛选:指出葱花的潜在作用
IF 1.8 Q3 PHARMACOLOGY & PHARMACY Pub Date : 2024-07-12 DOI: 10.4274/tjps.galenos.2023.88935
Özlem Oyardı, Mayram Hacıoğlu, Ebru Özdemir, Meryem Şeyda Erbay, Şükran Kültür, Çağla Bozkurt Güzel

Objectives: Plant extracts are important natural resources that may have antimicrobial and antibiofilm effects against pathogens. This study was conducted to investigate the in vitro antimicrobial activities of methanol extracts of some medicinal plants (Achillea nobilis subspecies neilreichii (A. Kern.) Velen., Aetheorhiza bulbosa (L.) Cass, Allium paniculatum L, Asphodelus aestivus Brot., Ballota nigra L., Cistus laurifolius L., Cistus salviifolius L., Dioscorea communis (L.) Caddick and Wilkin, Galium verum L., Hypericum triquetrifolium Turra, Paliurus spina-christi Mill., Primula vulgaris Huds. subspecies rubra (Sm.) Arcang., Ranunculus arvensis L. and Teucrium polium L.) from Balıkesir province in Türkiye.

Materials and methods: Preliminary antimicrobial activity screening was conducted for all extracts. Antibiofilm activity studies were conducted on mature Candida albicans biofilms. Moreover, the cytotoxicities of A. paniculatum flower extract on A549 and Vero cell lines were determined using a colorimetric tetrazolium-based assay.

Results: A. paniculatum flower, P. vulgaris root, C. laurifolius, C. salviifolius, and A. nobilis displayed good activity [minimum inhibitory concentrations (MIC): 9.75, 156, 312, 312 and 312 μg/mL, respectively] against C. albicans American Type Culture Collection 10231. Biofilm studies were conducted on these plant extracts. The methanol extract of A. paniculatum flower decreased the number of C. albicans [colony-forming unit (CFU)/mL] in mature biofilm statistically at 32 x MIC and higher concentrations (p < 0.01). A. paniculatum flower extract had a cytotoxic effect (killing more than 50% of cells) at high concentrations, and its effect on Vero cells was similar to that on A549 cells.

Conclusion: This study demonstrated the importance of the methanol extract of A. paniculatum flower as a natural alternative against C. albicans infections, including biofilms.

目的:植物提取物是一种重要的天然资源,可能对病原体具有抗菌和抗生物膜作用。本研究旨在研究一些药用植物(Achillea nobilis L.subsp.neilreichii(A.Kern.)Velen。,球叶Aetheohiza bulbosa(L.)Cass,葱(Allium paniculatum L),普通小麦(Aspodelus aestivus Brot.)。,Ballota nigra L.、Cistus laurifolius L.、Cistos salviifolius L.、Dioscorea consus(L.)Caddick&Wilkin、Galium verum L.、Hypericum triquetrifolium Turra、Palius spina christi Mill。,寻常报春花。亚种。rubra(Sm.)Arcing。,Ranunculus arvensis L.和Teucrium polium L.)。材料与方法:对所有提取物进行初步抗菌活性筛选。对白色念珠菌成熟生物膜进行了抗菌膜活性研究。此外,使用基于比色四氮唑的测定法测定了A.paniculatum花提取物对A549和Vero细胞系的细胞毒性。结果:A.paniculatum花、P.vulgaris根、C.laurifolius、C.salvifolius和A.nobilis对白色念珠菌ATCC 10231表现出良好的活性(MICs分别为9.75、156、312、312和312µg/ml)。对这些植物提取物进行了生物膜研究。在32xMIC及更高浓度下,圆锥花甲醇提取物可显著降低成熟生物膜中白色念珠菌的数量(cfu/ml)(P<0.01)。圆锥花甲醇提取液在高浓度下具有细胞毒性作用(杀死50%以上的细胞),对Vero细胞的作用与对A549细胞的作用相似。结论:本研究证明了圆锥花甲醇提取物作为包括生物膜在内的白色念珠菌感染的天然替代品的重要性。
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引用次数: 0
Olmesartan Medoxomil-Loaded Niosomal Gel for Buccal Delivery: Formulation, Optimization, and Ex Vivo Studies. 奥美沙坦酯-甲氧杂环己酯口腔给药胶:配方、优化和离体研究
IF 1.8 Q3 PHARMACOLOGY & PHARMACY Pub Date : 2024-07-12 DOI: 10.4274/tjps.galenos.2023.93765
Narahari Narayan Palei, Bibhash Chandra Mohanta, Jayaraman Rajangam, Prathap Madeswara Guptha

Objectives: Olmesartan medoxomil (OLM) is a low bioavailability antihypertensive drug. This study aimed to prepare and optimize an OLM niosomal gel and investigate drug permeation via a chicken buccal pouch.

Materials and methods: OLM-loaded niosome were prepared using a film hydration technique. The vesicle size, zeta potential, entrapment efficiency, and percentage cumulative drug release of niosome were evaluated. The niosomes were incorporated into a Carbopol 974P (1.5% w/v) gel, and the drug permeability of the niosomal gel was evaluated. The formulations of the niosomal gel were optimized using the Box-Behnken design. The optimized formulation was further characterized by transmission electron microscopy (TEM) and Fourier transform infrared radiation analysis.

Results: The particle size and zeta potential of the optimized niosomal formulations were 296.4 nm and -38.4 mV, respectively. Based on TEM analysis, the niosomes were found to be spherical in shape. The permeability, flux, and permeability coefficient of the optimized niosomal gel were 0.507 mg/cm2, 0.083 mg/cm2 × hour, and 041 cm/hour, respectively. Histopathological evaluation revealed that the niosomal gel had better permeability than the OLM gel.

Conclusion: Based on the results of the OLM niosomal gel, it can be concluded that the formulation can be beneficial in increasing bioavailability, resulting in better therapeutic efficacy.

目的:奥美沙坦酯是一种生物利用度低的降压药。本研究旨在制备和优化OLM niosomal凝胶,并研究药物通过鸡颊袋的渗透性。方法:采用膜水化技术制备OLM负载的羊膜。对囊泡大小、ζ电位、包封效率和药物累积释放百分比进行了评估。将niosomes掺入Carbopol 974P(1.5%w/v)凝胶中,并评估niosomes凝胶的药物渗透性。使用Box-Behnken设计优化了niosomal凝胶的配方。使用透射电子显微镜(TEM)和FTIR分析对优化的配方进行了进一步表征。结果:优化后的niobody制剂的粒径和ζ电位分别为296.4nm和-38.4mV。通过透射电镜分析,发现该囊泡呈球形。结果表明,优化后的niobody凝胶的渗透性、通量和渗透系数分别为0.507mg/cm2、0.083mg/cm2×h和041cm/h。组织病理学评估显示,与OLM凝胶相比,羊膜体凝胶具有更好的渗透性。结论:基于OLM羊膜凝胶的结果,可以得出结论,该制剂有利于提高生物利用度,从而产生更好的治疗效果。
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引用次数: 0
Evaluation of Herbal Products and Dietary Supplements Use in Patients with Respiratory Diseases Applied to Tertiary Health Institution. 三级医疗机构呼吸系统疾病患者使用草药产品和膳食补充剂的评价
IF 1.8 Q3 PHARMACOLOGY & PHARMACY Pub Date : 2024-07-12 DOI: 10.4274/tjps.galenos.2023.08504
Tuğba Subaş, Ufuk Özgen, Yılmaz Bülbül, Tevfik Özlü, Gülin Renda, Abdul Kadir Albayraktar

Objectives: In recent years, especially with the Coronavirus Disease of 2019 (COVID-19) pandemic, the use of herbal products for various health problems has been increasing worldwide. This study aimed to determine the frequency of herbal product/dietary supplement use, the most used products, and the factors affecting the use of these products in patients who applied to the Chest Diseases Clinic.

Materials and methods: This descriptive survey study was conducted at Chest Diseases Clinic using a face-to-face interview technique. Adult individuals with subacute respiratory complaints for > 3 weeks or a diagnosis of chronic chest disease were included in the study. The questionnaire form included questions about personal characteristics, data related to disease and treatment, use of herbal products/dietary supplements, and attitudes toward these products. A total of 444 participants with all the data included in the study. Descriptive statistics, chi-square, and binary logistic regression tests were used.

Results: It was determined that 49.3% of the participants used herbal products/dietary supplements, and the most frequently used products were honey, linden, ginger, lemon, and carob. According to the results of the binary logistic regression test, it was determined that patients over 60 years old [odds ratio (OR)= 2.0, 95% confidence interval (Cl): 1.1-3.8, p= 0.042], those with a high education level (OR= 2.0, 95% Cl: 1.1-3.6, p= 0.018), those who live in urban (OR= 1.8, 95% Cl: 1.1-3.0, p= 0.018), and those with a diagnosis of post-COVID syndrome (OR= 2.7, 95%, Cl: 1.3-5.5, p= 0.007) are more likely to use these products. It was determined that 57.9% of the participants used these products to relieve the symptoms of the disease.

Conclusion: Considering the high probability of using these products in patients with respiratory tract disease, it is essential for public health that health professionals question the use of these products and provide counseling on this issue.

目的:近年来,特别是随着新冠肺炎的流行,草药产品在全球范围内用于各种健康问题的情况不断增加。本研究旨在确定申请胸科疾病诊所的患者使用草药产品/膳食补充剂的频率、最常用的产品以及影响这些产品使用的因素。材料和方法:这项研究是一项描述性调查研究,在卡拉德尼兹工业大学法拉比医院胸科诊所采用面对面访谈技术进行。研究包括亚急性呼吸系统不适超过三周或诊断为慢性胸部疾病的成年个体。调查表包括关于个人特征、与疾病和治疗相关的数据、草药产品/膳食补充剂的使用以及对这些产品的态度的问题。444名没有遗漏数据的参与者被纳入研究。采用描述性统计、卡方检验和二元逻辑回归检验。结果:49.3%的参与者使用草药产品/膳食补充剂,最常用的产品是蜂蜜、菩提树、生姜、柠檬和角豆。根据二元逻辑回归检验的结果,确定60岁以上(OR=2.0,95%CI:1.1-3.8,p=0.042)、具有高教育水平(OR=2.0,95%Cl:1.1-3.6,p=0.018)、生活在城市(OR=1.8,95%CI:1.1-3.0,p=0.018,p=0.018和诊断为新冠肺炎后综合征(OR=2.7,95%,Cl:1.3-5.5,p=0.007)的患者更有可能使用这些产品。经测定,57.9%的参与者使用这些产品来缓解疾病症状。结论:考虑到这些产品在呼吸道疾病患者中使用的可能性很高,卫生专业人员对这些产品的使用提出质疑并就此问题提供咨询对公共卫生很重要。
{"title":"Evaluation of Herbal Products and Dietary Supplements Use in Patients with Respiratory Diseases Applied to Tertiary Health Institution.","authors":"Tuğba Subaş, Ufuk Özgen, Yılmaz Bülbül, Tevfik Özlü, Gülin Renda, Abdul Kadir Albayraktar","doi":"10.4274/tjps.galenos.2023.08504","DOIUrl":"10.4274/tjps.galenos.2023.08504","url":null,"abstract":"<p><strong>Objectives: </strong>In recent years, especially with the Coronavirus Disease of 2019 (COVID-19) pandemic, the use of herbal products for various health problems has been increasing worldwide. This study aimed to determine the frequency of herbal product/dietary supplement use, the most used products, and the factors affecting the use of these products in patients who applied to the Chest Diseases Clinic.</p><p><strong>Materials and methods: </strong>This descriptive survey study was conducted at Chest Diseases Clinic using a face-to-face interview technique. Adult individuals with subacute respiratory complaints for > 3 weeks or a diagnosis of chronic chest disease were included in the study. The questionnaire form included questions about personal characteristics, data related to disease and treatment, use of herbal products/dietary supplements, and attitudes toward these products. A total of 444 participants with all the data included in the study. Descriptive statistics, chi-square, and binary logistic regression tests were used.</p><p><strong>Results: </strong>It was determined that 49.3% of the participants used herbal products/dietary supplements, and the most frequently used products were honey, linden, ginger, lemon, and carob. According to the results of the binary logistic regression test, it was determined that patients over 60 years old [odds ratio (OR)= 2.0, 95% confidence interval (Cl): 1.1-3.8, <i>p</i>= 0.042], those with a high education level (OR= 2.0, 95% Cl: 1.1-3.6, <i>p</i>= 0.018), those who live in urban (OR= 1.8, 95% Cl: 1.1-3.0, <i>p</i>= 0.018), and those with a diagnosis of post-COVID syndrome (OR= 2.7, 95%, Cl: 1.3-5.5, <i>p</i>= 0.007) are more likely to use these products. It was determined that 57.9% of the participants used these products to relieve the symptoms of the disease.</p><p><strong>Conclusion: </strong>Considering the high probability of using these products in patients with respiratory tract disease, it is essential for public health that health professionals question the use of these products and provide counseling on this issue.</p>","PeriodicalId":23378,"journal":{"name":"Turkish Journal of Pharmaceutical Sciences","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2024-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47141937","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Characterization of Equilibrative Nucleoside Transport of the Pancreatic Cancer Cell Line: Panc-1. 胰腺癌细胞系Panc-1核苷平衡转运的表征
IF 1.8 Q3 PHARMACOLOGY & PHARMACY Pub Date : 2024-07-12 DOI: 10.4274/tjps.galenos.2023.86727
Sıla Appak Başköy, Amardeep Khunkhuna, Bianca Scuric, Zlatina Naydenova, Imogen R Coe

Objectives: Gemcitabine, a first-line chemotherapeutic nucleoside analog drug (NAD) for pancreatic cancer, faces limitations due to drug resistance. Characterizing pancreatic cancer cells' transport characteristics may help identify the mechanisms behind drug resistance, and develop more effective therapeutic strategies. Therefore, in this study, we aimed to determine the nucleoside transport properties of Panc-1 cells, one of the commonly used pancreatic adenocarcinoma cell lines.

Materials and methods: To assess the presence of equilibrative nucleoside transporter-1 (ENT-1) in Panc-1 cells, we performed immunofluorescence staining, western blot analysis, and S-(4-nitrobenzyl)-6-thioinosine (NBTI) binding assays. We also conducted standard uptake assays to measure the sodium-independent uptake of [3H]-labeled chloroadenosine, hypoxanthine, and uridine. In addition, we determined the half-maximal inhibitory concentration (IC50) of gemcitabine. Statistical analyses were performed using GraphPad Prism version 8.0 for Windows.

Results: The sodium-independent uptake of [3H]-labeled chloroadenosine, hypoxanthine, and uridine was measured using standard uptake assays, and the transport rates were determined as 111.1 ± 3.4 pmol/mg protein/10 s, 62.5 ± 4.8 pmol/mg protein/10 s, and 101.3 ± 2.5 pmol/mg protein/10 s, respectively. Furthermore, the presence of ENT-1 protein was confirmed using NBTI binding assays (Bmax 1.52 ± 0.1 pmol/mg protein; equilibrium dissociation constant 0.42 ± 0.1 nM). Immunofluorescence assays and western blot analysis also revealed ENT-1 in Panc-1 cells. The determined IC50 of gemcitabine in Panc-1 cells was 2 μM, indicating moderate sensitivity.

Conclusion: These results suggest that Panc-1 is a suitable preclinical cellular model for studying NAD transport properties and potential therapies in pancreatic cancer and pharmaceutical research.

目的:吉西他滨是一种核苷类似物药物,用于胰腺癌的一线化疗药物。细胞系的特征对于了解胰腺癌的耐药发展至关重要。在这项研究中,我们旨在确定Panc-1细胞的核苷转运特性,Panc-1细胞是常用的胰腺腺癌细胞系之一。材料和方法:为了评估Panc-1细胞中平衡核苷转运蛋白1 (ENT1)的存在,我们进行了免疫荧光染色、western blot分析和S-(4-硝基苄基)-6-硫代肌苷(NBTI)结合试验。我们还进行了标准摄取试验,以测量[3H]标记的氯腺苷、次黄嘌呤和尿苷的钠独立摄取。此外,我们测定了吉西他滨的半最大抑制浓度(IC50)。使用GraphPad Prism version 8.0 for Windows进行统计分析。结果:采用标准摄取法测定了[3H]标记的氯腺苷、次黄嘌呤和尿苷的钠独立摄取,其转运率分别为111.1±3.4 pmol/mg蛋白/10秒、62.5±4.8 pmol/mg蛋白/10秒和101.3±2.5 pmol/mg蛋白/10秒。此外,通过S-(4-硝基苄基)-6-硫代氨基甘氨酸(NBTI)结合实验证实了ENT1蛋白的存在(Bmax 1.52±0.1 pmol/mg蛋白;Kd 0.42±0.1 nM)。免疫荧光分析和western blot分析也显示在Panc-1细胞中存在ENT-1。测定的吉西他滨在Panc-1细胞中的IC50为2 μM,敏感性中等。结论:Panc-1是研究核苷类药物转运特性和潜在治疗胰腺癌和药物研究的合适临床前细胞模型。
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引用次数: 0
Proteomic Analysis of HepG2 Cells Reveals FAT10 and BAG2 Signaling Pathways Affected by a Protease Inhibitor from Tinospora cordifolia (Willd.) Hook. f. and Thoms Stem. Extract Among the Different Plant and Microbial Samples Analyzed. HepG2细胞的蛋白质组学分析揭示了不同植物和微生物样品中受Tinospora cordifolia茎提取物蛋白酶抑制剂影响的FAT10和BAG2信号通路
IF 1.8 Q3 PHARMACOLOGY & PHARMACY Pub Date : 2024-07-12 DOI: 10.4274/tjps.galenos.2023.75668
Bramhi Suresh Chougule, Kumar Gaurav, Mutthu Kumar, Nayana Mahadevaprasad, Nisarga Hosahalli Krishna, Sreya Lakshmi Ponnada, Somasekhara Derangula, Varalakshmi Kilingar Nadumane

Objectives: Dysregulation of proteolysis underlies diseases like cancer. Protease inhibitors (PIs) regulate many biological functions and hence have potential anticancer properties. With this background, the current study aimed to identify the PI from natural sources such as plants and microbes against trypsin (a protease), which was assayed against casein, using an ultraviolet spectrophotometer-based methodology.

Materials and methods: PI extracted from a few plants and microbial samples were screened for their PI activity against trypsin. The PI from the most promising source in our study, Tinospora cordifolia (Willd.) Hook. f. and Thoms. stem, was partially purified using ammonium sulfate precipitation followed by dialysis. The PI activity of the partially purified inhibitor was analyzed against chymotrypsin and collagenase enzymes, and the cytotoxic effect of the PI was checked on HepG2 (liver carcinoma) cells by MTT- [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide]- assay. Liquid Chromatograography Mass Spectrometry -based proteomic studies were performed on HepG2 cells to understand the signaling pathways affected by the PIs in the liver cancer cell line.

Results: Among the samples tested the PIs from T. cordifolia stem extract had the highest inhibitory activity (72.0%) against trypsin along with cytotoxicity to HepG2 cells. After partial purification by 80.0% ammonium sulfate precipitation, PI had increased inhibitory activity (83.0%) against trypsin and enhanced cytotoxicity (47.0%) to HepG2 cells. Proteomic analysis of the PI-treated HepG2 cells revealed that BAG2 and FAT10 signaling pathways were affected, which may have caused the inhibition of cancer cell proliferation.

Conclusion: PI from T. cordifolia stem has promising anticancer potential and hence can be used for further purification and characterization studies toward cancer drug development.

目的:蛋白质水解失调是癌症等疾病的基础。蛋白酶抑制剂调节多种生物功能,因此具有潜在的抗癌特性。在此背景下,本研究的重点是从植物和微生物等天然来源中鉴定抗胰蛋白酶(一种蛋白酶)的PI。
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引用次数: 0
Optimization of Enterocin Production from Probiotic Enterococcus faecium Using Taguchi Experimental Design. 田口实验设计优化益生菌粪肠球菌产肠毒素的研究
IF 1.8 Q3 PHARMACOLOGY & PHARMACY Pub Date : 2024-07-12 DOI: 10.4274/tjps.galenos.2023.83451
Dina Manny, Amr El-Waseif, Eslam Abd El-Wahed

Objectives: Enterocin is a significant broad-spectrum peptide antibiotic produced by Enterococcus faecium (E. faecium). Enterocin production by E. faecium was investigated using the Taguchi experimental design. The Taguchi models were used to save the time and effort required for optimizing the different conditions affecting its production. They were applied to optimize the conditions for enterocin production using the least number of experiments and the least number of required materials.

Materials and methods: Seven factors i.e., pH, temperature, time of incubation, aeration rate, inoculum size, carbohydrate concentration, and bile salt concentrations, each at three levels were selected and an orthogonal array layout of L273 was performed.

Results: The experimental results indicated that the best incubation conditions were; 48 hours incubation on a nutrient medium at pH 6.5, temperature at 25 °C, aeration rate at 0 round per minute, inoculum size 20 mL, and bile salt concentration. It was 5%, and the carbon concentration was 2.0%. All these factors combined led to the best enterocin production by E. faecium.

Conclusion: This optimization of enterocin production by the Taguchi experimental models emphasized some important results regarding the interaction of the different driving factors leading to the best enterocin production in one experiment.

肠毒素是由粪肠球菌产生的一种重要的广谱肽类抗生素。采用田口实验设计研究了粪肠球菌产肠毒素的情况。使用田口模型是为了节省优化影响其生产的不同条件所需的时间和精力。它们被应用于通过最少的实验次数和最少的所需材料量来优化肠肽生产的条件。选择三个水平的七个因素,即pH、温度、培养时间、通气率、接种物大小、碳水化合物浓度和胆盐浓度,并进行L27(3)的正交阵列布局。实验结果表明,最佳培养条件为:;在pH 6.5、温度25°C、通气速度0 rpm、接种量20 ml、胆汁盐浓度的营养培养基上培养48小时。为5%,碳浓度为2.0%。所有这些因素加在一起导致粪肠球菌产生最佳的肠毒素。田口实验模型对肠毒素生产的优化强调了关于不同驱动因素相互作用的一些重要结果,这些驱动因素在一个实验中产生了最佳的肠毒素生产。
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引用次数: 0
The Effect of Sucrose and Yeast Extract on Total Phenolic, Flavonoid, and Anthocyanin of Lactic-Acid-Fermented Mangosteen Fruit Peel (Garcinia mangostana L.). 蔗糖和酵母提取物对乳酸发酵山竹果皮总酚、类黄酮和花青素的影响
IF 1.8 Q3 PHARMACOLOGY & PHARMACY Pub Date : 2024-07-12 DOI: 10.4274/tjps.galenos.2023.91267
Komang Dian Aditya Putra, G A Desya Pradnyaswari, Putu Sanna Yustiantara, I Made Agus Gelgel Wirasuta, Eka Indra Setyawan

Objectives: This study aimed to determine the most suitable concentration of sucrose and yeast extract (SYE) and its impact on the levels of total phenol, flavonoid, and anthocyanin (TPFA) for lactic acid fermentation in mangosteen fruit peel.

Materials and methods: In this study, the primary components were mangosteen fruit peel, SYE, and lactic acid bacteria starter. The experimental design was conducted using the Factorial Design method. The colorimetric method was used to determine the total phenol (Folin-Ciocalteu reagent) and total flavonoid (AlCl3 reagent). In addition, the differential pH method was used to determine the total anthocyanins using KCl and the CH3COONa reagent.

Results: The addition of SYE during the fermentation of mangosteen fruit peel significantly increased the concentrations of TPFA compared with the control (p value of 0.0001). The high sucrose concentration and low yeast extract produced the highest TPFA levels in mangosteen rind fermentation.

Conclusion: The use of SYE affects the levels of TPFA in lactic acid-fermented mangosteen fruit peel, with the most suitable concentrations obtained using sucrose (45 g/L) and yeast extract (2.5 g/L).

目的:本研究旨在确定山竹果皮乳酸发酵(LAF)的最佳蔗糖和酵母提取物(SYE)浓度及其对总酚、类黄酮和花青素(TPFA)水平的影响。材料与方法:本研究以山竹果皮、蔗糖、酵母提取物、乳酸菌发酵剂为主要成分。实验设计采用因子设计法。采用比色法测定总酚(Folin-Ciocalteu试剂)和总黄酮(AlCl3试剂)。此外,用KCl和CH3COONa试剂,用差示pH法测定总花青素。结果:与对照组相比,在山竹果皮发酵过程中添加SYE可以显著提高TPFA的浓度(p值为0.0001)。在山竹皮发酵中,高蔗糖浓度和低酵母提取物产生的TPFA水平最高。结论:SYE的使用影响乳酸发酵山竹果皮中TPFA的水平,蔗糖(45g/L)和酵母提取物(2.5g/L)的浓度最为合适。
{"title":"The Effect of Sucrose and Yeast Extract on Total Phenolic, Flavonoid, and Anthocyanin of Lactic-Acid-Fermented Mangosteen Fruit Peel (<i>Garcinia mangostana</i> L.).","authors":"Komang Dian Aditya Putra, G A Desya Pradnyaswari, Putu Sanna Yustiantara, I Made Agus Gelgel Wirasuta, Eka Indra Setyawan","doi":"10.4274/tjps.galenos.2023.91267","DOIUrl":"10.4274/tjps.galenos.2023.91267","url":null,"abstract":"<p><strong>Objectives: </strong>This study aimed to determine the most suitable concentration of sucrose and yeast extract (SYE) and its impact on the levels of total phenol, flavonoid, and anthocyanin (TPFA) for lactic acid fermentation in mangosteen fruit peel.</p><p><strong>Materials and methods: </strong>In this study, the primary components were mangosteen fruit peel, SYE, and lactic acid bacteria starter. The experimental design was conducted using the Factorial Design method. The colorimetric method was used to determine the total phenol (Folin-Ciocalteu reagent) and total flavonoid (AlCl<sub>3</sub> reagent). In addition, the differential pH method was used to determine the total anthocyanins using KCl and the CH<sub>3</sub>COONa reagent.</p><p><strong>Results: </strong>The addition of SYE during the fermentation of mangosteen fruit peel significantly increased the concentrations of TPFA compared with the control (<i>p</i> value of 0.0001). The high sucrose concentration and low yeast extract produced the highest TPFA levels in mangosteen rind fermentation.</p><p><strong>Conclusion: </strong>The use of SYE affects the levels of TPFA in lactic acid-fermented mangosteen fruit peel, with the most suitable concentrations obtained using sucrose (45 g/L) and yeast extract (2.5 g/L).</p>","PeriodicalId":23378,"journal":{"name":"Turkish Journal of Pharmaceutical Sciences","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2024-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44710121","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
GC-MS Profiling and Pharmacological Potential of Physconia venusta (Ach.) Poelt. 金星藻(Physconia venusta)Poelt的气相色谱-质谱分析及其药理潜力
IF 1.8 Q3 PHARMACOLOGY & PHARMACY Pub Date : 2024-07-12 DOI: 10.4274/tjps.galenos.2023.91126
Ibtissem Zeghina, Ibtissem El Ouar, Maya Abir Tartouga, Mohamed Badreddine Mokhtari, Daniel Elieh-Ali-Komi, Lynda Gali, Chawki Bensouici

Objectives: Lichens are complex symbiotic organisms that generate various bioactive compounds with significant therapeutic value. We investigated the chemical composition and bioactivity of the acetone extract of the Algerian lichen Physconia venusta (Ach.) poet.

Materials and methods: Phytochemical screening was performed using gas chromatography-mass spectrometry (GC-MS). The antibacterial activity was assessed against Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis, Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes, and Bacillus subtilis using an agar diffusion test with the determination of the minimal inhibition concentration (MIC), while the antioxidant activity was determined using different chemical methods (DPPH, ABTS, CUPRAC, reducing power, superoxide anion scavenging, β-carotene bleaching, and metal chelate). In addition, cytotoxic activity was tested using Artemia salina (Brine shrimp) bioassay.

Results: The studied extract exhibited intense antibacterial activity against E. coli and S. aureus with inhibition diameters of 28 ± 0.01 and 22 ± 0.01 mm, respectively, with a MIC value of 6.25 mg/mL and a selectivity index of 2.8. The obtained extract showed different antioxidant trends depending on the selected assay. GC-MS analysis revealed many secondary metabolites.

Conclusion: P. venusta, a type of lichen, is a potential source of bioactive substances that could be used in pharmaceuticals.

目的:地衣是一种复杂的共生生物,能产生各种具有重要治疗价值的生物活性化合物。我们试图研究阿尔及利亚地衣Physconia venusta(Ach.)Poelt的丙酮提取物的化学成分和生物活性。材料与方法:采用气相色谱-质谱联用技术进行植物化学筛选。使用琼脂扩散试验测定最小抑制浓度(MIC),评估对大肠杆菌、铜绿假单胞菌、肠炎沙门氏菌、斑疹伤寒杆菌、金黄色葡萄球菌、李斯特菌和枯草芽孢杆菌的抗菌活性,同时使用不同的化学方法(DPPH、ABTS、CURAC、还原力、超氧化物阴离子清除、β-胡萝卜素漂白和金属螯合物)测定抗氧化活性。此外,使用卤虫(盐水虾)生物测定法测试细胞毒性活性。结果:该提取物对大肠杆菌和金黄色葡萄球菌具有较强的抗菌活性,抑菌直径分别为28±0.01和22±0.01mm,MIC值为6.25mg/ml,选择性指数为2.8。所获得的提取物根据所选择的测定显示出不同的抗氧化趋势。GC-MS分析显示存在许多次级代谢产物。结论:金星苔草是一种潜在的药用生物活性物质来源。
{"title":"GC-MS Profiling and Pharmacological Potential of <i>Physconia venusta</i> (Ach.) Poelt.","authors":"Ibtissem Zeghina, Ibtissem El Ouar, Maya Abir Tartouga, Mohamed Badreddine Mokhtari, Daniel Elieh-Ali-Komi, Lynda Gali, Chawki Bensouici","doi":"10.4274/tjps.galenos.2023.91126","DOIUrl":"10.4274/tjps.galenos.2023.91126","url":null,"abstract":"<p><strong>Objectives: </strong>Lichens are complex symbiotic organisms that generate various bioactive compounds with significant therapeutic value. We investigated the chemical composition and bioactivity of the acetone extract of the Algerian lichen <i>Physconia venusta</i> (Ach.) poet.</p><p><strong>Materials and methods: </strong>Phytochemical screening was performed using gas chromatography-mass spectrometry (GC-MS). The antibacterial activity was assessed against <i>Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis, Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes</i>, and <i>Bacillus subtilis</i> using an agar diffusion test with the determination of the minimal inhibition concentration (MIC), while the antioxidant activity was determined using different chemical methods (DPPH, ABTS, CUPRAC, reducing power, superoxide anion scavenging, β-carotene bleaching, and metal chelate). In addition, cytotoxic activity was tested using <i>Artemia salina</i> (Brine shrimp) bioassay.</p><p><strong>Results: </strong>The studied extract exhibited intense antibacterial activity against <i>E. coli</i> and <i>S. aureus</i> with inhibition diameters of 28 ± 0.01 and 22 ± 0.01 mm, respectively, with a MIC value of 6.25 mg/mL and a selectivity index of 2.8. The obtained extract showed different antioxidant trends depending on the selected assay. GC-MS analysis revealed many secondary metabolites.</p><p><strong>Conclusion: </strong><i>P. venusta</i>, a type of lichen, is a potential source of bioactive substances that could be used in pharmaceuticals.</p>","PeriodicalId":23378,"journal":{"name":"Turkish Journal of Pharmaceutical Sciences","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2024-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49472622","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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Turkish Journal of Pharmaceutical Sciences
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