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   Table of Contents - Current issue
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May-June 2022
Volume 17 | Issue 3
Page Nos. 231-333

Online since Monday, April 18, 2022

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ORIGINAL ARTICLES  

A simple LC-MS/MS method for pharmacokinetic study of carvedilol and 4/-hydroxyphenyl carvedilol at a low dose p. 231
Wanna Eiamart, Nantaporn Prompila, Yaowatree Jumroen, Nonlanee Sayankuldilok, Pajaree Chariyavilaskul, Supeecha Wittayalertpanya
DOI:10.4103/1735-5362.343077  
Background and purpose: The study was aimed at validating a simple, rapid, and low-cost LC-MS/MS method for carvedilol and 4/-hydroxyphenyl carvedilol assay in human plasma. The validated method was applied to investigate the pharmacokinetics after a low dose of 6.25 mg. carvedilol. Experimental approach: In this study, the plasma was extracted by liquid-liquid extraction and evaporated the organic layer to dryness, then both analytes in the residue were reconstituted and detected by LC- MS/MS. The method was validated following the guideline on bioanalytical method validation. Thirty-one healthy volunteers participated in the pharmacokinetic study. After 10 h of fasting, each volunteer received one tablet of 6.25 mg carvedilol orally. Blood samples were collected at 16 prescheduled time points. The plasma samples were analyzed for pharmacokinetics. Findings/Results: The method was linear over a range of 0.050-50.049 ng/mL for carvedilol and 0.050-10.017 ng/mL for 4/-hydroxyphenyl carvedilol. Crucial validated results reached the requirements of selectivity, accuracy, precision, and stability. Pharmacokinetics of carvedilol and 4/-hydroxyphenyl carvedilol were evaluated which showed Cmax at 21.26 ± 9.23 and 2.42 ± 2.07 ng/mL; AUC0-t 66.95 ± 29.45 and 5.93 ± 3.51 ng.h/mL; AUC0-inf 68.54 ± 30.11 and 6.78 ± 3.49 ng.h/mL; and T1/2 6.30 ± 1.95 and 6.31 ± 6.45 h, respectively. Conclusion and implications: The validated method was able to detect and quantify both analytes in plasma samples and can be applied to the pharmacokinetic study of carvedilol and 4/-hydroxyphenyl carvedilol after receiving carvedilol at 6.25 mg orally.
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Investigation of the relationship between miR-33a, miR-122, erythrocyte membrane fatty acids profile, and serum lipids with components of metabolic syndrome in type 2 diabetic patients p. 242
Fatemeh Masoudi, Mohammad Reza Sharifi, Morteza Pourfarzam
DOI:10.4103/1735-5362.343078  
Background and purpose: MicroRNAs (miRNAs) are small non-coding RNA molecules acting as critical regulators of post-transcriptional gene expression. MiR-33a and miR-122 have a crucial role in cholesterol and lipid metabolism. Therefore, their dysregulation may contribute to metabolic abnormality and their inhibition may be a useful therapeutic strategy. The objective of the present study was to investigate the relationship between miR-33a, miR-122, erythrocyte membrane fatty acids profile, and serum lipids with components of metabolic syndrome in an Iranian population suffering from type 2 diabetes mellitus (T2DM). Experimental approach: Expression of miR-33a and miR-122 was measured by real-time polymerase chain reaction and erythrocyte membrane fatty acid profiles were analyzed by gas chromatography-mass spectrometry. Findings/Results: T2DM patients with and without metabolic syndrome had significantly higher miR-33a and miR-122 levels compared to controls. MiRNAs were significantly correlated with saturated fatty acid (SFAs), total SFAs/total polyunsaturated fatty acids (PUFAs) ratio, fasting plasma glucose, triacylglycerols, insulin and homeostatic model assessment of insulin resistance. In addition, there was a significant negative correlation between miR-33a and miR-122 levels and PUFAs, total PUFAs/total SFAs ratio and omega 6 fatty acids. Conclusion and implications: Considering the roles of miR-33a and miR-122 in cholesterol and lipids metabolism, it may be concluded that the measurement of their expression may be useful as a potential additional biomarker for cardiometabolic derangement in T2DM patients. In addition, these findings may suggest that the inhibition of these miRNAs by anti-miRNA therapies may be explored as a potential therapeutic strategy.
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Synthesis, characterization, molecular docking, antimalarial, and antiproliferative activities of benzyloxy-4-oxopyridin benzoate derivatives p. 252
Marjan Mohebi, Neda Fayazi, Somayeh Esmaeili, Mahboubeh Rostami, Fereshteh Bagheri, Alireza Aliabadi, Parvin Asadi, Lotfollah Saghaie
DOI:10.4103/1735-5362.343079  
Background and purpose: Malaria and cancer are two major health issues affecting millions of lives annually. Maltol complexes and derivatives have been extensively investigated as chemotherapeutic and antimalarial activities. In this study, the design, synthesis, biological activities, and docking study of a novel series of pyridinones derivatives were reported. Experimental approach: The chemical structures of synthesized compounds were approved by FTIR, 1HNMR, 13CNMR, and mass spectroscopies. The antimalarial activity was evaluated through β-hematin inhibition assay and the cytotoxicity activities were evaluated against PC12 and fibroblast cell lines via MTT and cell uptake assays. To theoretically investigate the ability of compounds to inhibit hemozoin formation, the synthesized compounds were docked in a heme sheet to explore their binding mode and possible interactions. Findings/Results: β-Hematin inhibition assay showed acceptable activity for 7f, 7c, and 7d compounds and the molecular docking study showed 7h and 7f had effective interactions with the heme sheet. The cytotoxic study revealed compound 4b (IC50 = 18 μM) was significantly more active against PC12 cells than docetaxel (IC50 = 280 μM). The observations of cell uptake images were also shown both cell penetration and monitoring potential of synthesized compounds. Conclusion and implications: The compounds showed a moderate ability to inhibition of heme polymerization and also good interaction with heme through molecular docking was observed. Additionally, some of them have a good cytotoxic effect on the study2 cell line. So further study on these compounds can lead to compounds that can be considered as anti-malarial and/or anticancer agents.
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Hydroethanolic extract of Lavandula angustifolia ameliorates vincristine-induced peripheral neuropathy in rats p. 265
Shamim Sahranavard, Mona Khoramjouy, Mahsima Khakpash, Seyyed Ahmad Askari, Mehrdad Faizi, Mahmoud Mosaddegh
DOI:10.4103/1735-5362.343080  
Background and purpose: Peripheral neuropathy is one of the most common adverse effects of cancer chemotherapy. Vincristine is prescribed to treat a variety of carcinomas, including lymphoma and leukemia, and may cause progressive peripheral neuropathy due to the damage of microtubules and mitochondria of neurons and affects inflammatory processes. This study was designed to evaluate the effects of Lavandula angustifolia hydroalcoholic extract (LHE) of aerial part on vincristine-induced peripheral neuropathy in a rat model. Experimental approach: Neuropathy was induced in rats by daily intraperitoneal administration of vincristine (0.1 mg/kg for 2 weeks). Following the induction of neuropathy, animals were treated with the LHE (100, 200, and 400 mg/kg, p.o.) or pregabalin (20 mg/kg, IP) for 2 weeks, and their responses to vincristine-induced hyperalgesia and locomotor impairment were measured. Findings/Results: LHE, at the dose of 400 mg/kg, showed analgesic effects in response to thermal hyperalgesia, tactile allodynia, and gait impairment. Also, pregabalin (20 mg/kg, IP) improved the symptoms of vincristine-induced peripheral neuropathy. Conclusions and implications: According to the results, we can conclude that LHE alleviates neuropathic symptoms of vincristine and the effect is probably related to the presence of phenols and flavonoids in the extract.
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Policies and measures implemented to reduce the impact of international sanctions on Iran's pharmaceutical sector p. 274
Farideh Emami, Abdol Majid Cheraghali
DOI:10.4103/1735-5362.343081  
Background and purpose: Continuous evaluation of the policies and interventions could explore their efficacies in improving the accessibility and availability of medicines in the local market. This study explained the health system policies of the Islamic Republic of Iran concerning the impact of the economic sanctions on the local pharmaceutical market and addressed the issue of whether these policies were able to improve patients’ access to medicines. Experimental approach: In this study, qualitative and quantitative research methods were used. In the qualitative part, semi-structured interviews with pharmaceutical system experts were used. In the next step, the structural analysis method was used. In the quantitative part, numerical sales data of the selected medicines were extracted and analyzed. Findings/Results: Reported statistics regarding the presence of the medicines in the market indicates the obvious fluctuations in the numerical sales of medicines in the studied years. This may indicate that the policies implemented are not able to fully compensate for the negative effects of sanctions and improve access to medicine. In addition, according to some experts, policy, and management weaknesses are mainly rooted in unresolved domestic hurdles which have exacerbated the effects of international sanctions on the country’s pharmaceutical market. Conclusions and implications: Effective policy-making in response to economic sanctions can reduce the negative effect of international sanctions and result in drug shortages. Results of this study showed despite efforts made by the Iran health sector to subside the impact of sanctions on the pharmaceutical sector, there is an obvious disruption of the medicine supply chain in the market.
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Design and production of a novel chimeric human growth hormone superagonist fused to human Fc domain p. 284
Samira Mirbaha, Marzieh Rezaei, Rahman Emamzadeh, Sayyed Hamid Zarkesh Esfahani
DOI:10.4103/1735-5362.343082  
Background and purpose: Growth hormone (GH) has been known as a crucial metabolic hormone expressed at the pituitary and the other number of cells and tissues and responsible for body growth. Because of the short half-life of GH, daily subcutaneous injections were shown to be more effective for GH therapy. This represents a burden for patients. So, there is a strong effort from the industry to create a long-acting form of GH and lots of technologies like GH fusion proteins are used to increase GH half-life. Experimental approach: In this study, the Fc domain of human IgG1 with serine-glycine linkers was attached to the C-terminal of a GH superagonist via molecular cloning. The presence of recombinant vector in E. coli host was confirmed by PCR. SDS-PAGE and western blot analysis showed the expression of recombinant proteins in the bacterial lysate. The binding ability to growth hormone receptors is determined by ELISA. Findings / Results: Our results showed that the novel SupGH-Fc has a good binding affinity to its receptor in ELISA in comparison to standard GH, although it has a big size. Conclusion and implications: Our data in this study clearly demonstrated the expression of the SupGH-Fc in a recombinant protein expression system. It is an introduction to the production of the new recombinant GH, which can bind to its receptor more effectively than commercial growth hormones and also might have a longer half-life.
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Alantolactone and ZnO nanoparticles induce apoptosis activity of cisplatin in an ovarian cancer cell line (SKOV3) p. 294
Shahriar Alipour, Ghader Babaei, Shiva Gholizadeh-Ghaleh Aziz, Somayeh Abolhasani
DOI:10.4103/1735-5362.343083  
Background and purpose: Ovarian cancer is one of the leading causes of cancer mortality in women. Despite the increase in cases of this cancer, the current therapeutic strategy is not effective. This study aimed to investigate the effect of cisplatin (Cis) with alantolactone (ALT) and ZnO nanoparticles (ZnONPs) in inducing apoptosis in SKOV3 ovarian cancer cells line. Experimental approach: To evaluate the viability of SKOV3 cells and determine the IC50 of Cis, ALT, and ZnONPs, MTT assay was used. Real-time PCR and western blotting were used to evaluate the expression levels of genes (XIAP, cyclin D1, Bcl-2, Bax, and MDM2) and proteins (XIAP, cyclin D1, Bcl-2, Bax), respectively. Also, cellular ROS levels were assessed by fluorimetry. Findings / Results: Our results showed that ALT and ZnONPs significantly increased the response to Cis in SKOV3 cells compared to the control and this response is remarkably increased in the triple combination (ALT-Cis-ZnONPs). The expression of XIAP, cyclin D1, and Bcl-2 genes and proteins in the groups treated with ALT, Cis, and ZnONPs as a single agent, double and triple combination were significantly reduced compared to the control, while Bax was generally shown an increase. Also, the level of intracellular ROS is higher in the treatment groups than in the control group and the highest increase was observed in the triple combination. Conclusion and implications: Taken together, our data demonstrated the potential therapeutic approach of using ALT and ZnONPs that may enhance the apoptotic effects of Cis on the SKOV3 cells.
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Further evidence to support acute and chronic anti-inflammatory effects of Nasturtium officinale p. 305
Mostafa Mostafazadeh, Heibatollah Sadeghi, Hossein Sadeghi, Vahid Zarezade, Abolghasem Hadinia, Esmaeel Panahi Kokhdan
DOI:10.4103/1735-5362.343084  
Background and purpose: Previously, we reported the anti-inflammatory properties of Nasturtium officinale (watercress) in several models of acute inflammation. This study was designed to explore the effects of topical and systemic administrations of N. officinale in the two chronic inflammatory models and to evaluate the role of TNF-α and IL-1β in these effects. Experimental approach: Folin-Ciocalteu and aluminum chloride methods were used to estimate the extract’s total phenol and flavonoid content, respectively. Carrageenan-induced paw edema was carried out and TNF-α and IL-1β concentrations in the carrageenan-treated paw tissue were determined. Formalin injection into rat hind paws (7 days) and the application of 12-O-tetradecanoyl phorbol-13-acetate (TPA) on mouse ears (9 days) were used to simulate chronic inflammation. Furthermore, a histological assessment of the inflamed tissues was carried out. Findings/Results: The extract’s flavonoid and phenolic contents were 90.26 ± 4.81 mg rutin equivalents/g and 68 ± 8.16 gallic acid equivalents/g gallic acid, respectively. N. officinale pretreatment in all doses administered considerably decreased carrageenan-induced edema. The extract also reduced IL-1β levels in carrageenan- treated paws while did not affect TNF-α levels. Oral and topical administrations of N. officinale considerably reserved the paw and ear edema. The extract also ameliorated the tissue injuries due to formalin and TPA challenges. Conclusion and implications: The data confirmed the topical and systemic anti-inflammatory effects of watercress against two chronic models of inflammation. They suggested that these properties are not related to TNF-α but could be attributed to inhibition of IL-1β and inhibition of leukocyte infiltration.
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Comparison of follicular T helper cells, monocytes, and T cells priming between newly diagnosed and rituximab-treated MS patients and healthy controls p. 315
Sheida Yahyazadeh, Nafiseh Esmaeil, Vahid Shaygannejad, Omid Mirmosayyeb
DOI:10.4103/1735-5362.343085  
Background and purpose: The use of anti-CD20 monoclonal antibodies like rituximab (RTX) to deplete B cells has practical therapeutic implications in multiple sclerosis (MS) patients. However, the therapy’s impact on other immune cells is also important. Therefore, in this study, we assessed the effects of RTX therapy on Tfh cells, T cells, T cells priming, and monocytes in MS patients compared to newly-diagnosed MS patients and healthy subjects. Experimental approach: Thirty newly-diagnosed and RTX-treated MS patients and healthy control were included. Peripheral blood mononuclear cells were isolated from whole blood for assessment of Tfh cells, CD4+, CD8+, CD4+CD45RA+, CD3+HLA-DR+, and CD3+CD4+CD25+ T cells by flow cytometry. Whole blood was lysed by lysis solution to assess CD45+CD14+ monocytes by flow cytometry. Also, the serum level of interleukin 21 was measured by the ELISA method. Findings / Results: We showed that RTX treatment led to a decrease in Tfh cells and their predominant cytokine, interleukin 21. Also, we found a statistically significant reduction in CD3+HLA-DR+ and CD3+CD4+CD25+ T cells in RTX-treated patients compared to new cases and healthy control. Moreover, we found a decrease in the CD45+ CD14+ monocyte population in the RTX-treated group compared to the healthy control. Conclusion and implications: Our data suggest that following treatment with RTX, Tfh cells, monocytes, and T cells priming declined happened, and fewer T cells were activated. Also, due to the interaction between B cells and Tfh cells, Tfh targeting could be assessed as a therapeutic strategy in MS.
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Beneficial effects of kebar grass (Biophytum petersianum klotzsch) ethanol extract to increase motor reflex and spatial memory in mice offspring (Mus musculus) from lactating mothers exposed to carbofuran p. 324
Epy Muhammad Luqman, Eka Pramyrtha Hestianah, Widjiati Widjiati, Suryo Kuncorojakti, Viski Fitri Hendrawan
DOI:10.4103/1735-5362.343086  
Background and purpose: This study aimed to determine the potency of kebar grass ethanol extract to overcome an increase in cerebellar neuronal cell necrosis, which has an impact on decreasing motor reflex function and spatial memory of mice from lactating mothers exposed to carbofuran. Experimental approach: Forty lactating mice were divided into four groups, 10 each; including control, T1 (carbofuran 0.0125 mg/day), T2 (vitamin C 5 mg + carbofuran 0.0125 mg/day), T3 (kebar grass extract 3.375 mg + carbofuran 0.0125 mg/day). The mice were orally administered with carbofuran, vitamin C, and kebar grass extract on days 0 to 14 postnatal. On the 15th day, brains of the mice were necropsied to measure the levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH), H&E staining; motor reflex tests were performed on 10-day-old mice, and the mice aged 30 days were tested on their swimming and spatial memory. Findings / Results: Carbofuran caused an increase in MDA, GSH, neuronal cell necrosis, surface righting reflex, a decrease in SOD, swimming ability, and spatial memory. Kebar grass extract and vitamin C administration decreased MDA, GSH, neuron necrosis, surface righting reflex, and increased SOD, swimming ability, and spatial memory. Conclusion and implications: Exposing to carbofuran in lactating mice caused brain oxidative stress, impaired motor reflexes, and spatial memory in mice offspring. Kebar grass extract and vitamin C administration prevented brain oxidative stress and inhibited disorders in motor reflexes, and spatial memory in mice offspring. Kebar grass extract administration was more effective than vitamin C.
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