فهرست مطالب

Advanced Pharmaceutical Bulletin
Volume:8 Issue: 3, Aug 2018

  • تاریخ انتشار: 1397/07/07
  • تعداد عناوین: 20
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  • Arun Kumar, Arun Nanda *, Sandeep Kumar Pages 355-363
    Pharmaceutical Co-crystals are not new, they have gained much attention since the last decade among scientists and pharmaceutical industry. Pharmaceutical co-crystals are multicomponent systems composed of two or more molecules and held together by non-covalent interactions. The development of pharmaceutical co-crystals, a new solid crystalline form, offer superior physico-chemical properties (such as melting point, stability, solubility, permeability, bioavailability, taste masking, etc.) without altering the pharmacological properties. Recently, with the upsurge in the growth of Pharmaceutical co-crystals, the major concern is over the regulatory status of co-crystals. With the new guidelines from United States Food and Drug Administration (USFDA) and European Medicines Agency (EMA), the status has become even more complicated due to significantly different opinions. This review highlights whether co-crystals fulfil the requirements for the grant of a patent or not and how cocrystals are going to affect the present scenario of pharmaceuticals
    Keywords: Pharmaceutical Co-crystals, Crystal Engineering, Regulatory guidelines, Patents
  • Velid Unsal * Pages 365-376
    Heavy metals taken into the organism can make the toxic effects on the metabolism in various ways. For example, they may interact with proteins to alter and inhibit their enzymatic and structural functions. Mercury is one of the toxic elements that are widely distributed in nature. Mercury toxicity poses a serious threat to human health. It is an element that causes oxidative stress to increase in individuals, leading to tissue damage. Oxidative stress is the result of the imbalance between the production of oxidative species and cellular antioxidant defense. Phytotherapy continues to play an important role in health care. Natural phytotherapeutic antioxidants, exhibit a broad sequence of biological impacts, including anti-oxidative stress, anti-aging, anti-toxicicity and anticancer. Many studies have also shown that the phytotherapeutic agents play an important role in the removal of mercury from the tissue and in reducing oxidative stress. Our goal in this review was to investigate alternative ways of extracting the mercury in the tissue.
    Keywords: Mercury, Antioxidants, Phytotherapy, Oxidative stress
  • Taher Entezari, Maleki *, Sajad Khiali, Afshin Gharekhani Pages 377-382
    Purpose
    Isotretinoin is the most effective anti-acne drug with a long-term remission. However; it contains severe teratogenic effects with serious adverse drug reactions, which limits the use of medication.
    Methods
    To review the use of isotretinoin during pregnancy, we carried out a comprehensive search of literature in Google Scholar, Scopus and PubMed/Medline from their inception until April 2015.
    Results
    Database searching identified 277 records, of which, 38 articles were retrieved according to abstract and title assessment. After full-text review, 17 articles were excluded and finally, a total of 21 studies met the inclusion criteria. Data showed an increased pattern in the use of isotretinoin. In some studies, health care providers were not fully adhered to the risk reduction programs in pregnancy. Exposing to isotretinoin among pregnant women has still occurred due to detrimental adherence to risk reduction programs which resulted in live-born infants with different kinds of abnormalities.
    Conclusion
    Despite the known serious adverse effect of isotretinoin, the use of drug was not based on the guidelines in some cases, which needs more attentions to prevent the severe drug related problems.
    Keywords: Isotretinoin, Acne, Utilization pattern, Teratogenic effects, Pregnancy
  • Fatemeh Atyabi, Mehdi Yousefi, Dariush Shanehbandi, Alireza Mohajjel Nayebi, Mohammad, Sadegh Soltani Zangbar, Farhad Jadidi, Niaragh, Leili Aghebati, Maleki, Jafar Majidi, Reza Jafari, Naime Majidi Zolbanin Pages 383-393
    Purpose
    Targeted treatment of breast cancer through combination of chemotherapeutic agents and siRNA had been drawing much attention in recent researches. This study was carried out to evaluate mucin1 aptamer-conjugated chitosan nanoparticles containing docetaxel and cMET siRNA on SKBR3 cells.
    Methods
    Nano-drugs were characterized by transmission electron microscope, Zetasizer and loading efficiency calculation. siRNA entrapment onto nanoparticles, stability of siRNA-loaded nanoparticles and conjugation of mucin1 aptamer to nanoparticles were evaluated via separate electrophoresis. Cellular uptake of the targeted nanoparticles was evaluated through GFP-plasmid expression in mucin1+ SKBR3 vs. mucin1- CHO cells. Protein expression, cell viability and gene expression were assessed by Western Blotting, MTT assay, and Quantitative Real Time-PCR, respectively.
    Results
    Characterization of nano-drugs represented the ideal size (110.5± 3.9 nm), zeta potential (11.6± 0.8 mV), and loading efficiency of 90.7% and 88.3% for siRNA and docetaxel, respectively. Different gel electrophoresis affirmed the conjugation of aptamers to nanoparticles and entrapment of siRNA onto nanoparticles. Increased cellular uptake of aptamer-conjugated nanoparticles was confirmed by GFP expression. cMET gene silencing was confirmed by Western Blotting. The significant (p ≤0.0001) impact of combination targeted therapy vs. control on cell viability was shown. Results of Quantitative Real Time-PCR represented a remarkably decreased (p ≤0.0001) expression of the studied genes involving in tumorigenicity, metastasis, invasion, and angiogenesis (STAT3, IL8, MMP2, MMP9, and VEGF) by targeted combination treatment vs. control.
    Conclusion
    The mucin1 aptamer-conjugated chitosan nanoparticles, containing docetaxel and cMET siRNA, is suggested for treatment of mucin1+ metastatic breast cancer cells. However, further studies should be conducted on animal models.
    Keywords: Aptamer, Chitosan, cMET siRNA, Docetaxel, Metastatic breast cancer
  • Faezeh Alihosseini, Sepideh Arbabi Bidgoli, Setareh Haghighat, Ahoo Afshar Nasab, Seyyedeh Elaheh Mousavi, Solmaz Ghaffari *, Nooshin Kianvash, Seyed Mahdi Rezayat Sorkhabadi Pages 395-400
    Purpose
    Wound healing is a natural biologic process, but the duration of it may take too long. Trying to shorten this process is one of the challenges for scientists. Many technologies were applied to achieve this goal as well as nanotechnology. In this study semi solid formulations containing curcumin and ampicillin solid lipid nanoparticles (SLNs) were prepared to evaluate as burn wound healing agent.
    Methods
    Curcumin as an anti-inflammatory and anti-bacterial agent and ampicillin as an antibiotic were applied. In-vitro and in-vivo evaluations were carried out. Particle size, loading efficiency, release profile, morphology and anti-bacterial efficacy of desired nanoparticles were evaluated at first. Then the remaining of the antibacterial effect in semi solid preparations was studied. Animal studies for both toxicology using rabbits and skin burn model using rats were designed. Pathology studies after applying of formulations was done too.
    Results
    Desired nanoparticles were spherical in shape and particle size in range of 112-121 nm, with low zeta potential. For increasing stability of particles they were freeze dried using cryoprotectant. Lyophilized particles show no significant size enlargement. Results showed that both ointment and gel preparations have reasonable anti-bacterial effects, both of them cause increasing in the rate of wound healing in comparison with placebos and control groups and none of the formulations showed acute toxicity.
    Conclusion
    It seems that using nanotechnology could shorten wound healing process to reduce treatment costs and increase compliance of patients.
    Keywords: Wound healing, Solid Lipid Nanoparticles, In-vivo, Semi solids
  • Jessada Prasomkij, Kamon Panrat, Wiwat Pichayakorn, Jirapornchai Suksaeree * Pages 401-410
    Purpose
    The objective of the present investigation was to prepare and evaluate transdermal patches for nicotine.
    Methods
    Pectin isolated from the hulls of Monthong durian or leaves of Krueo Ma Noy was used as a matrix membrane for the controlled release of nicotine and compared with commercial pectin. The mechanical properties, moisture uptake, and Fourier transform infrared spectra were characterized. The in vitro stability of these patches was evaluated and compared to commercial nicotine patches.
    Results
    The mechanical properties of the patches made from isolated pectin were greater than those prepared from commercial pectin; brittle commercial patches were obtained after nicotine loading. The moisture uptake of the patches made with isolated pectin was in the range of 30.20-44.29%. There was no incompatibility between the ingredients of the nicotine transdermal patches or any degradation of the drug. The matrix layer made from isolated pectin controlled the nicotine release more effectively than did commercial nicotine patches. In addition, these patches were stable at in a refrigerator (approximately 4±2 °C) and at ambient temperature (approximately 30±2 °C) for 3 months, retaining 90% of the loaded nicotine.
    Conclusion
    Our study suggests that using isolated pectin as the matrix layer should control the release of nicotine from transdermal patches.
    Keywords: Isolated pectin, Nicotine transdermal patches, Durian fruit, Cissampelos pareira, Krueo Ma Noy
  • Zahra Sobhani *, Marziyeh Zare, Soliman Mohammadi Samani Pages 411-417
    Purpose
    Due to limited oral bioavailability of doxorubicin (Dox) many efforts during the last decades focused on the development of novel delivery systems to overcome these limitations. In the present study, Dox encapsulated chitosan nanoparticles were prepared to evaluate the intestinal permeation of Dox via oral administration.
    Methods
    Nanoparticles were fabricated based on ionic gelation method using tripolyphosphate. Some physicochemical properties, such as nanoparticle size and morphology, loading efficiency and in vitro drug release in 3 different pH values (5.0, 6.8 & 7.4) were evaluated. Intestinal permeations of free Dox and Dox loaded in nanoparticles were assessed using rat intestinal sac model.
    Results
    The nanoparticles were spherical shape with average size of 150 10 nm. The entrapment and loading efficiency of Dox were up to 40% and 23%, respectively. According to the release profiles, up to 30% of loaded drug was released within 6hrs and the remaining amount of Dox was released more gradually, but this pattern was related to pH of the medium. The amount of drug released at acidic condition (pH 5.0) was greater than other pHs. The intestinal permeation of Dox increased nearly up to 90% by loading in chitosan nanoparticles.
    Conclusion
    Using chitosan nanoparticles presents a potential safe drug delivery system for oral administration of Dox. In vivo studies and the determined pharmacokinetic and pharmacodynamic of Dox loaded chitosan nanoparticles after oral administration are planned for future studies.
    Keywords: Doxorubicin, Chitosan nanoparticles, Oral delivery, Intestinal permeation, Everted rat gut method
  • Rajeswari Surya Anusha Venna, Srikanth Reddy Poreddy, Venkata Subbaiah Kotakadi, Saritha Damineni, Subhash Chandra Bose Penjuri *, Nagaraju Ravouru Pages 419-427
    Purpose
    The main aim of the present investigation was to enhance the solubility of poorly soluble Gliclazide by nanocrystallization.
    Methods
    In present investigation gliclazide nanocrystals were prepared by sonoprecipitation using Pluronic F68, Poly Vinyl Alcohol (PVA), Poly ethylene Glycol 6000 (PEG), Poly Vinyl Pyrrolidine (PVP K30) and Sodium Lauryl Sulphate (SLS) as stabilizers. Fourier Transform Infrared Spectroscopic study (FTIR), Differential Scanning Calorimetry (DSC) and X ray diffraction (XRD) studies were conducted to study the drug interactions. Size and zeta potential of the nanocrystals were evaluated. In vitro and in vivo studies of nanocrystals were conducted in comparison to pure gliclazide.
    Results
    The Gliclazide nanocrystals (GN) showed mean particle size of 131±7.7 nm with a zeta potential of -26.6 mV. Stable nanocrystals were formed with 0.5% of PEG 6000. FTIR, DSC and XRD studies of nanocrystals showed absence of interactions and polymorphism. SEM photographs showed a change in morphology of crystals from rod to irregular shape. There is an increase in the saturation solubility and the percentage drug release from formulation GN5 (Optimized Gliclazide Nanocrystals) was found to be 98.5 in 15 min. In the in vivo study, GN5 nanocrystals have reduced the blood glucose level to 296.4±4.26 mg/dl in 12 hr. The nanocrystals showed lower tmax and higher Cmax values as compared to pure gliclazide.
    Conclusion
    The prepared nanocrystals of gliclazide were stable without any drug polymer interactions. Increase in the dissolution of nanocrystals compared to pure gliclazide and significant reduction in blood glucose level in vivo indicated better bioavailability of the nanocrystals. Therefore, it is concluded that nanocrystal technology can be a promising tool to improve solubility and hence dissolution of a hydrophobic drug.
    Keywords: Dissolution rate, Gliclazide, Particle size, Sonoprecipitation, Zeta potential
  • Masoud Dadkhah, Hadi Feizi *, Alireza Abdanipour, Mohsen Alipour Pages 429-435
    Purpose
    The antiapoptotic effect of ghrelin in various cell lines including bone marrow stromal cells (BMSCs) has been proved. However, the real mechanism of this effect is not clear. Caspase3 and Bcl2 are well-known pro- and antiapoptotic regulatory genes in eukaryotes. The aim of the study was to find out the effect of ghrelin on Caspase 3 and Bcl2 change in BMSCs.
    Methods
    Rat BMSCs were cultivated in DMEM. Passage 3 BMSCs were treated with ghrelin 100 μM for 48 h. Real-time PCR for Caspase 3 and Bcl2 was carried out from B (untreated BMSCs), BH (BMSCs treated with 125 µM H2O2), BGH (BMSCs treated with 100 µM ghrelin then 125 µM H2O2) and BG (BMSCs treated with 100 µM ghrelin) groups. For immunofluorescence, cells were incubated with anti Caspase 3 and Bcl2monoclonal antibodies. Primary antibodies were visualized using the FITC method. All data are presented as means ± SEM. Values of P<0.05 were considered statistically significant.
    Results
    Ghrelin decreased mRNA expressions of Caspase-3 significantly as compared to the BH group (P<0.05). Also, Bcl-2 gene expression showed an increment in BG group as compare with BH and BGH groups (P<0.05). A high present of Bcl-2 positive cells were observed in the BGH group while Caspase-3 positive cells were significantly decreased in the BGH group compared with the BH group (P<0.05).
    Conclusion
    Ghrelin probably enhances BMSCs viability through regulation of pro- and antiapoptotic genes Caspase 3 and Bcl2. However the signaling pathway of this effect should be elucidated in the future.
    Keywords: Ghrelin, Caspase 3, Bcl2, H2O2, Rat, BMSCs
  • Maryam Hemmatzadeh, Hamed Mohammadi, Farhad Babaie, Behzad Baradaran *, Dariush Shanehbandi, Behzad Mansoori, Mehrdad Ebrazeh, Mehdi Yousefi Pages 437-445
    Purpose
    Snail-1 is a transcription factor, which takes part in EMT, a process related to the emergence of invasion and cancer progression. The purpose of this study was to evaluate the effect of Snail-1 silencing on the human esophageal squamous cell carcinoma cell line, namely TE-8, in vitro.
    Methods
    In this study, transfection of Snail-1 specific siRNA was conducted into TE-8 cells. The relative mRNA expression levels of Snail-1, Vimentin, CXCR4 and MMP-9 and transcription levels of miR-34a and let-7a were investigated by quantitative Real-time PCR. Western blotting was carried out to evaluate the Snail-1 protein level. Migration assay of TE-8 cells was also performed following the presence or absence of Snail-1 specific siRNA. MTT and TUNEL assays were performed to evaluate cell viability after Snail-1 silencing.
    Results
    It was found that treatment of cancer cells with the Snail-specific siRNA effectively downregulated the expression of Snail-1 in both mRNA and protein levels, and vimentin, CXCR4, and MMP-9 in mRNA level. However, it elevated the transcript levels of miR-34a and let-7a expressions. Furthermore, transfection of cancer cells with the Snail-specific siRNA significantly induced apoptosis in TE8 cells. Moreover, suppression of Snail-1 led to diminished cell migration.
    Conclusion
    It seems that Snail-specific siRNA can significantly interrupt esophageal cancer cell migration and reduce metastatic-related factors and induce miR-34a and let-7a in vitro. The bottom line is that therapeutic approaches via targeting Snail-1 can be used for ESCC treatment, suggesting that other possible target molecules for ESCC therapy require to be explored.
    Keywords: Esophageal cancer, Snail-1, siRNA, Apoptosis, Metastasis
  • Seyed Ali Hosseinian, Aliakbar Haddad Mashadrizeh, Samaneh Dolatabadi * Pages 447-455
    Purpose

    Epithelial cell adhesion molecule (EpCAM) is a dominant antigen in human colon carcinoma tissue. Topology features of this antigen are different in normal and malignant conditions; for instance, EpCAM is much less accessible to antibodies in normal cells than in cancerous tissues. Hence, EpCAM has been considered as a suitable candidate for cancer target therapy via immunotoxins (ITs) development. In this study, attention was focused on the stability assessment of anti-EpCAM-IT (anti-Ep-IT) to design a novel IT.

    Methods

    The 3D structures of the antibody template and the toxin segment of anti-Ep-IT were retrieved from PDB. Discovery Studio3.0 was used to separate the ligands and water molecules. The antibody (Ab) fragment of anti-Ep-IT was aligned using protein blast (BLAST-p), and SWISS-MODEL database was used for Ab modeling. IT modeling was accomplished using MODELLER 9.15. Also, GROMACS 5.07 was used for molecular dynamic (MD) simulation step. Moreover, ERRAT and RAMPAGE databases were used for quality assessment of the structures.

    Results

    BLAST-p results indicated that antibody moiety of IT has the highest E-value and query coverage scores to the monoclonal antibody (mAb) 4D5MOC-B. Modeling by SWISS-MODEL provided a reasonable template for Ab portion compared to MODELLER. The best modeled full-length IT with the lowest RMSD values was selected. Finally, RMSD plot for MD stage demonstrated constant values from 7000ps to 20000ps.

    Conclusion

    In general, both modeling results and their quality evaluations were satisfactory for designing IT. Moreover, RMSD plot revealed that IT stability was preserved during the simulation. Overall, our findings led to modeling and simulation of the anti-Ep-IT with more structural stability.

    Keywords: EpCAM, Cancer therapy, Immunotoxin, Simulation, Stability
  • Azadeh Montaseri *, Jafar Soleimani Rad, Mohammad Karimipour, Nahid Karimian Fathi, Raheleh Farahzadi, Maryam Eyvazi Pages 457-464
    Purpose
    Application of Mummy material for treatment of different diseases such as bone fracture, cutaneous wounds and joint inflammation has been advised since hundred years ago in Persian traditional medicine. Due to the claims of indigenous people and advice of traditional medicine for application of this material in healing of bone fractures, this study has been designed to evaluate whether Mummy material can promote the differentiation of mesenchymal stem cells into osteoblasts and enhance the expression of bone specific genes and proteins.
    Methods
    Adipose derived stem cells (ASCs) at fourth cell passage were divided into control, osteogenesis group (received osteogenic medium), Mummy group (received Mummy at concentration of 500 µg/ml). ASCs in the fourth group were treated with both osteogenic medium and Mummy (500µg/ml). Cells in all groups were harvested on days 7, 14 and 21 days for further evaluation through Real time RT-PCR, Von kossa staining, Immunocytochemistry and flowcytometery.
    Results
    Treatment of ASCs with Mummy at concentration of 500µg/ml promotes the expression level of Osteocalcin, RUNX-2 and β1-integrin genes in different time points but that of the Osterix did not changed. Furthermore the expression of Osteocalcin protein enhanced significantly in ASCs treated with Mummy detected by Immunocytochemistry and flowcytometery technique compared to the control groups. The results of this study also showed that treatment of ASCs with Mummy resulted in formation of mineral deposits which was evaluated by Von Kossa staining method.
    Conclusion
    Obtained data from this study reveals that Mummy is a potent enhancer for differentiation of ASCs into osteoblasts in in vitro system, probably through increasing the level of bone specific genes and proteins.
    Keywords: Osteoblast, Adipose derived stem cells, Differentiation, Transcription factor
  • Hamed Bashiri, Mehryar Habibi Roudkenar *, Mohammad Ali Jalili, Yoshikazu Kuwahara, Masoud Hamidi, Amaneh Mohammadi Roushandeh, Ali Hosseini, Fatemeh Amiri Pages 465-470
    Purpose
    Poor survival rate of mesenchymal stem cells (MSCs) following their transplantation is one of the major challenges in their therapeutic application. Therefore, it is necessary to augment the viability of the MSCs in order to improve their therapeutic efficacy. Several strategies have been used to overcome this problem. Preconditioning of MSCs with oxidative stresses has gained a lot of attention. Therefore, in the present study, we investigated the effects of simultaneous preconditioning of MSCs with hydrogen peroxide and serum deprivation stresses on their survival and resistance to stressful conditions.
    Methods
    MSCs were isolated from human umbilical cord blood. To perform simultaneous preconditioning, the cells were cultured in DMEM medium containing 1, 2.5 and 5 percent FBS and different concentrations of H2O2 (5, 10, 15, 20, 25, 30, 35, 40, 50, 60, 80 and 100 µM) for 24 hrs. Then, the cells were cultured in recovery culture medium. Finally, one group of the cells was exposed to a lethal concentration of H2O2 (300µM), and the other cells were cultivated in FBS free DMEM medium as the lethal situation. In addition, the percentage of apoptotic cells was analyzed using Caspase 3 assay kit.
    Results
    Simultaneous preconditioning of the MSCs with 15µM H2O2 plus serum deprivation, 2.5% FBS, significantly increased the resistance of the cells to the toxicity induced following their cultivation in FBS free DMEM medium. It exerted the protective effect on the cells after treating with the lethal dose of H2O2 as well.
    Conclusion
    Simultaneous preconditioning of MSCs with oxidative and serum deprivation stresses enhances their survival against harsh conditions, which might increase the viability and stability of the MSCs following their transplantation.
    Keywords: Hydrogen peroxide, Mesenchymal Stem Cells, Serum deprivation, Simultaneous preconditioning, Survival, Harsh microenvironments
  • Farzaneh Foroughinia, Bahram Movahed Nouri, Javad Kojuri, Mohammad Ali Ostovan * Pages 471-478
    Purpose

    Studies have revealed that patients with chronic kidney disease (CKD) are more susceptible to adverse effects of percutaneous coronary intervention (PCI). In addition, the role of elevated high sensitive C-reactive protein (hs-CRP) in the prediction of adverse cardiac outcomes after coronary stent implantation has already been shown. Therefore, in this study, we aimed to evaluate the effect of omega-3 supplementation on hs-CRP and 30-day major adverse cardiac events (MACE) in patients with CKD undergoing elective PCI.

    Methods

    In this randomized trial, 80 CKD patients who were candidates for elective PCI, were randomly assigned to two groups; the first group received a single dose of omega-3 (2500 mg, 12 h before PCI) as well as the standard drug regimen of PCI and the second group received placebo plus the standard therapy (325 mg loading dose of aspirin, 600 mg loading dose of clopidogrel, and weight-adjusted intravenous heparin). Hs-CRP levels were measured at baseline and 24 h after the intervention as a primary endpoint. The secondary endpoint was the incidence of MACE over a 30-day period after PCI.

    Results

    Omega-3 did not significantly decrease post-PCI serum level of hs-CRP; however, the overall 30-day MACE was significantly lower in the omega-3 group compared to the control group (p=0.05).

    Conclusion

    Our results revealed the positive effect of the omega-3 supplement on decreasing 30-day MACE; hence, omega-3 may be considered as an effective adjunctive therapy to the standard drug regimen used before PCI. The evaluation of the effect of omega-3 on long-term MACE is recommended for future studies.

    Keywords: Chronic kidney disease, High sensitive C-reactive protein, Major adverse cardiac events, Omega-3 supplement, Percutaneous coronary intervention
  • Laleh Payahoo, Alireza Ostadrahimi *, Yaser Kahjebishak, Mohammad Asghari Jafarabadi Pages 479-487
    Purpose
    Obesity as a serious public health problem worldwide, results in the incidence of many chronic diseases. Obesity has been recognized as a chronic low-grade inflammation disorder. Altered endocannabinoid system tone is also involved in the pathogenesis of obesity. The present study aimed to investigate the effects of oleoylethanolamide supplementation on inflammatory biomarkers and oxidative stress in obese people.
    Methods
    This randomized, double-blind, placebo-controlled clinical trial was carried out on 60 healthy obese people in 2016 in Tabriz, Iran. Eligible subjects were randomly divided into intervention (received daily, two 125 mg OEA capsules) and control groups (the same amounts of starch) and treated for 8 weeks. Blood samples (5 ml) were taken in fasting state at the baseline and at the end of the study. The concentrations of MDA and TAS were measured using a spectrophotometer. A high sensitive-C reactive protein level was measured by Immunoturbidimetry assay using the commercial kit. IL-6 and TNF-α levels were assayed by the ELISA method. The differences between groups were assessed by ANCOVA and statistical significance was determined at p<0.05.
    Results
    Analysis was done on 56 participants who continued intervention until the end of the study. A significant decrease in the IL-6 and TNF-α serum concentrations was observed in the intervention group (p<0.001). Changes in other variables were undetectable (p>0.05).
    Conclusion
    The use of OEA as a complementary pharmacotherapy agent could be effective in improving inflammation and oxidative stress in obese people. Future studies are needed to confirm the obtained results.
    Keywords: Inflammation, Endocannabinoids, Obesity, Oleoylethanolamide, Oxidative stress
  • Amir Hooshang Mohammadpour , Saeed Nazemi , Fatemeh Mashhadi , Atefeh Rezapour , Mohammad Afshar , Sepideh Afzalnia , Afsaneh Mohammadi , Hamid Reza Mashreghi Moghadam , Maryam Moradian , Seyed Mohammad Hasan Moallem , Saeed Falahaty , Azadeh Zayerzadeh , Sepideh Elyas* Pages 489-493
    Purpose
    Coronary artery calcification (CAC) is utilized as an important tool for global risk assessment of cardiovascular events in individuals with intermediate risk. Ecto phosphodiesterase/nucleotide phosphohydrolase-1(ENPP1) converts extracellular nucleotides into inorganic pyrophosphate and it is a key regulator of tissue calcification that adjusts calcification in tissues like vascular smooth muscle cells. The main purpose of this clinical study was to find out the correlation between ENPP1 serum concentration and CAC in human for the first time.
    Methods
    In this study 83 patients (16 diabetic patients and 67 non-diabetic patients) with coronary artery disease who fulfilled inclusion and exclusion criteria, entered the study. For all patients a questionnaire consisting demographic data and traditional cardiovascular risk factors were completed. Computed tomography (CT)-Angiography was carried out to determine coronary artery calcium score and enzyme-linked immunosorbent assay (ELISA) method was used for measuring ENPP1 serum concentrations.
    Results
    There was a reverse significant correlation between ENPP1 serum concentration and total CAC score and also CAC of right coronary artery (RCA) (P<0.05) in non-diabetic patients.
    Conclusion
    On the basis of our results, ENPP1 serum concentration may be a suitable biomarker for coronary artery disease at least in non-diabetic patients. However, more studies with higher sample size are necessary for its confirmation.
    Keywords: Coronary Artery Calcification, ENPP1, Biomarker, Inorganic pyrophosphate, Glycoprotein 1- nucleotides
  • Yogesh Joshi, Srinath Muppalaneni, Alborz Omidian, David Jude Mastropietro, Hamid Omidian * Pages 495-505
    Purpose
    The purpose of this study was to determine the effects of thermal processing and antioxidant formulation variables on the abuse deterrence performance of a high molecular weight poly(ethylene oxide) (PEO) polymer.
    Methods
    A 24 factorial design with one categorical factor (antioxidant type) and three continuous factors (curing time, curing temperature, % antioxidant) was used. Abuse deterrence performance was evaluated using solution viscosity, surface melting temperature, and mechanical strength. Thermal degradation of PEO powders before compaction was also studied using DSC, FTIR spectroscopy, and viscosity analysis.
    Results
    Our results showed that curing temperature and type of antioxidant can significantly affect the deterrence performance of PEO. The main effect plot for viscosity shows the most prominent factors affecting viscosity are curing temperature and type of antioxidant. However, curvature in the linear model obtained was not sufficient to completely describe the behavior. For surface melting temperature, butylated hydroxytoluene was associated with higher surface melting temperatures compared to ascorbic acid. Additionally, higher percent of antioxidant resulted in higher melting temperature. Particle size distribution to indicate mechanical strength showed no significant effects of tested factors. This suggests that comminution method has more prominent effect on tablet fragment size than the formulation and processing factors studied.
    Conclusion
    While heat confers the mechanical strength to the polymer, it can diminish its physical stability and solution state viscosity. The experimental studies showed that prolonged exposure to high temperatures, even in the presence of antioxidants, can severely hamper polymer deterrence performance in both solid and solution states.
    Keywords: Abuse deterrent formulation, Injecting drug use, Crush resistance, Opioid abuse, Poly(ethylene oxide)
  • Riris Istighfari Jenie, Sri Handayani, Ratna Asmah Susidarti, Linar Zalinar Udin, Edy Meiyanto Pages 507-516
    Purpose
    Breast cancer cells with overexpression of HER2 are known to be more aggressive, invasive, and resistant to chemotherapeutic agent. Brazilin, the major compound in the Caesalpinia sappan L. (CS) heartwood, has been studied for it's anticancer activity. The purpose of this study was to investigate the cytotoxic and antimigratory activity of brazilin (Bi) in combination with doxorubicin (Dox) on MCF-7/HER2 cells.
    Methods
    Cytotoxic activities of Bi individually and in combination with Dox were examined by MTT assay. Synergistic effects were analyzed by combination index (CI). Apoptosis and cell cycle profiles were observed by using flow cytometry. Migrating and invading cells were observed by using a Boyden chamber assay. Levels of MMP2 and MMP9 activity were observed by using a gelatin zymography assay. Levels of HER2, Bcl-2, Rac1, and p120 protein expression were observed by using an immunoblotting assay.
    Results
    The results of the MTT assay showed that Bi inhibited MCF-7/HER2 cell growth in a dose-dependent manner with an IC50 of 54 ± 3.7 µM. Furthermore, the combination of Bi and Dox showed a synergistic effect (CI <1). Flow cytometric analysis of Bi and its combination with Dox showed cellular accumulation in the G2/M phase and induction of apoptosis through suppression of Bcl-2 protein expression. In the Boyden chamber assay, gelatin zymography, and subsequent immunoblotting assay, the combination Bi and Dox inhibited migration, possibly through downregulation of MMP9, MMP2, HER2, Rac1, and p120 protein expression.
    Conclusion
    We conclude that Bi enhanced cytotoxic activity of Dox and inhibited migration of MCF-7/HER2 cells. Therefore, we believe that it has strong potential to be developed for the treatment of metastatic breast cancer with HER2 overexpression.
    Keywords: Brazilin, Doxorubicin, Cytotoxic effect, Migration, MCF-7, HER2 cells
  • Sandro Rostelato Ferreira *_Cháriston André Dal Belo_Pedro Ismael da Silva Junior_Stephen Hyslop_Léa Rodrigues Simioni_Thomaz Augusto Alves Rocha e Silva Pages 517-522
    Purpose

    Rhinella schneideri is a toad found in many regions of the South America. The poison of the glands has cardiotoxic effect in animals and neuromuscular effects in mice and avian preparation. The purpose of this work was to identify the toxin responsible for the neuromuscular effect in avian and mice neuromuscular preparation.

    Methods

    The methanolic extract from R. schneideri poison was fractioned by reversed phase HPLC. The purity and molecular mass were determined by LC/MS mass spectrometry. Chick biventer cervicis and mouse phrenic-nerve diaphragm were used as neuromuscular preparations to identify the toxin.

    Results

    The purification resulted in 32 fractions, which 4 of them were active in neuromuscular preparation. The toxin of fraction 20 were chosen for better reproducibility of the whole extract activity and its molecular mass was 730.6 Da. The toxin produced facilitation of the muscle contraction followed by a complete neuromuscular blockade in chick biventer cervicis preparation in 90 min without interfering with the exogenous response to ACh and KCl. The quantal content was increased from 128 ± 13 (control) to 216 ± 44 (after 5 min and sustained until 60 min) in the presence of the toxin.

    Conclusion

    In conclusion, our results demonstrated that the neuromuscular action of the poison of Rhinella schneideri is a multitoxin effect. More, the present work first isolated a 730.6 Da toxin that better represent the whole poison neuromuscular effect, to which is attributed a presynaptic action in avian and mouse neuromuscular preparation.

    Keywords: Neurotransmitter release, Presynaptic toxin, Neuromuscular junction, Isolated fraction
  • Farouk Kamel El, Baz, Dalia Osama Saleh *, Gehad Abdel Raheem Abdel Jaleel, Rehab Ali Hussein Pages 523-528
    Purpose
    Aging is associated with hepatic morphological and physiological deterioration due to the accumulation of endogenous and exogenous free radicals and the resultant oxidative stress. The present study aims to investigate the effect of Haematococcus pluvialis microalgae on hepatic changes associated with D-galactose (D-Gal)-induced aging in rats.
    Methods
    Aging was induced in rats by daily intraperitoneal injection of D-Gal (200 mg/kg/day) for eight consecutive weeks. D-Gal-injected rats were treated by astaxanthin (ATX)-rich H. pluvialis biomass, its carotenoid and polar fractions for two weeks. Twenty four hours after the last dose, blood samples were collected and the liver tissues were isolated for further biochemical and histopathological examinations.
    Results
    D-Gal induced aging was associated with an elevation in serum liver function parameters, hepatic oxidative stress biomarkers viz., catalase (CAT), glutathione transferase (GST) and myeloperoxidase (MPO), as well as decreased expression of nuclear factor like-2 (Nrf2). Moreover, induction of aging exhibited an elevation of hepatic inflammatory cytokine; interleukin-6 (IL-6) and its modulator; nuclear factor Kappa B (NF-KB). However, treatment of D-Gal injected rats with ATX-rich H. pluvialis restored the serum liver function parameters as well as hepatic CAT, GST and MPO levels with an elevated expression of Nrf2. Treatment with ATX-rich H. pluvialis was also accompanied with a decrease in hepatic levels of NF-KB and IL-6. Histopathological examination emphasized all the previous results. Similarly, all trans-astaxanthin showed high affinity towards Nrf2 with -7.93 kcal/mol estimated free energy of binding as well as moderate affinities towards IL-6 and NF-KB through a docking study.
    Conclusion
    ATX-rich H. pluvialis showed beneficial effects by ameliorating the hepatic changes associated with D-Gal induced aging in rats due to its modulatory role of the Nrf2/Keap pathway.
    Keywords: Hepatic modulation, D-galactose, Aging, Haematococcus pluvialis, Astaxanthin