In Vitro and in Vivo Enhancement of Antitumoral Activity of Liposomal Antisense Oligonucleotides by Cineole as a Chemical Penetration Enhancer

Cellular uptake and cytoplasmic release of liposomal antisense oligonucleotides (AsODNs), which can act as rate-limiting steps, are still remained to be completely optimized. Here, the possibility of enhancing such processes at cellular and animal levels by cineole, as a penetration enhancer, was investigated. A cationic nanoliposome containing an AsODN against PKC-α and a cineole-containing nanoliposome were prepared and characterized. The effect of nanoliposomal cineole on sequence-specific cytotoxicity of nanoliposomal AsODN against A549, was studied in vitro (MTT, flow cytometry, fluorescence microscopy, and real time PCR) and in vivo (xenograft lung tumor in nude mice) using different concentrations and treatment times. Results showed specific cytotoxicity of nanoliposomal AsODN was increased significantly from 11 to 25 when A549 cells were exposed to 10 μg/mL cineole for 1 or 4 hours. This inhibitory effect was further increased to about 40 when the concentration was increased to 40 μg/mL for 1 hour. In animal studies, cineole significantly decreased the tumor volume (about 75) and increased its doubling time from 13 days to 31 days. A linear relationship exists between cineole concentration and its enhancement effects. Finally it was concluded that cineole, and possibly other membrane fluidizers, can improve nanoliposomal gene therapy at cellular and animal levels. © 2015 Hamid Reza Moghimi et al

Design, modeling, expression, and chemoselective PEGylation of a new nanosize cysteine analog of erythropoietin

Reza Ahangari Cohan1, Armin Madadkar-Sobhani2,3, Hossein Khanahmad1, Farzin Roohvand4, Mohammad Reza Aghasadeghi4, Mohammad Hossein Hedayati5, Zahra Barghi5, Mehdi Shafiee Ardestani4, Davoud Nouri Inanlou1, Dariush Norouzian11Research and Development Department, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran; 2Department of Bioinformatics, Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran; 3Department of Life Sciences, Barcelona Supercomputing Center, Barcelona, Spain; 4Hepatitis and AIDS Department, Pasteur Institute of Iran, Tehran, Iran; 5Quality Control Department, Production and Research Complex, Pasteur Institute of Iran, Tehran, IranBackground: Recombinant human erythropoietin (rhEPO) is considered to be one of the most pivotal pharmaceutical drugs in the market because of its clinical application in the treatment of anemia-associated disorders worldwide. However, like other therapeutic proteins, it does not have suitable pharmacokinetic properties for it to be administrated at least two to three times per week. Chemoselective cysteine PEGylation, employing molecular dynamics and graphics in in silico studies, can be considered to overcome such a problem.Methods: A special kind of EPO analog was elicited based on a literature review, homology modeling, molecular dynamic simulation, and factors affecting the PEGylation reaction. Then, cDNA of the selected analog was generated by site-directed mutagenesis and subsequently cloned into the expression vector. The construct was transfected to Chinese hamster ovary/dhfr- cells, and highly expressed clones were selected via methotrexate amplification. Ion-immobilized affinity and size exclusion (SE) chromatography techniques were used to purify the expressed analog. Thereafter, chemoselective PEGylation was performed and a nanosize PEGylated EPO was obtained through dialysis. The in vitro biologic assay and in vivo pharmacokinetic parameters were studied. Finally, E31C analog Fourier transform infrared, analytical SE-high-performance liquid chromatography, zeta potential, and size before and after PEGylation were characterized.Results: The findings indicate that a novel nanosize EPO31-PEG has a five-fold longer terminal half-life in rats with similar biologic activity compared with unmodified rhEPO in proliferation cell assay. The results also show that EPO31-PEG size and charge versus unmodified protein was increased in a nanospectrum, and this may be one criterion of EPO biologic potency enhancement.Discussion: This kind of novel engineered nanosize PEGylated EPO has remarkable advantages over rhEPO.Keywords: nanoPEGylated EPO, cysteine PEGylation, pharmacokinetic propert

Global, regional, and national burden of colorectal cancer and its risk factors, 1990–2019: a systematic analysis for the Global Burden of Disease Study 2019

Funding: F Carvalho and E Fernandes acknowledge support from Fundação para a Ciência e a Tecnologia, I.P. (FCT), in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy i4HB; FCT/MCTES through the project UIDB/50006/2020. J Conde acknowledges the European Research Council Starting Grant (ERC-StG-2019-848325). V M Costa acknowledges the grant SFRH/BHD/110001/2015, received by Portuguese national funds through Fundação para a Ciência e Tecnologia (FCT), IP, under the Norma Transitória DL57/2016/CP1334/CT0006.proofepub_ahead_of_prin

The Effect of Using Patency File on Apical Transportation in Canals Prepared with Passive Step Back Technique

Background: The purpose of this study was to assess the effect of patency file on apical transportation in curved canals prepared with passive step back (P.S.B) technique. Methods: This is an interceptive experimental, Invitro, study. Human extracted first permanent molars with 19-23mm length and curvature of 15-35 degrees (Schneider method) were considered for use in this study. Simple sampling was done and 35 teeth for each group was selected. In experimental group A canal preparation was done with P.S.B technique and usage of a # 10 file (as Patency file) between successive files. In experimental group B preparation was done in the same way except for usage of patency file. Pre and post operative radiographs were taken with similar condition. Radiographs were scanned and changes in canal curvature was determined using four different methods [Schneider, Weine, Long – Axis techniques (L. A.T), and Digital image overlay technique (overlay. T)] , using Idrisi for windows and AutoCAD softwares. Results: The mean of canal transportation angle in experimental group A with Schneider, Weine , LAT, and Over Lay techniques was: 7.006 ± 3.478, 12. 285 ± 6.032, 4.376 ± 3.516, 3.147 ± 2.744 respectively. Mean of canal transportation angle in experimental group B with the same methods was also determined: 8.009 ± 4.178, 13.55 ± 7.602, 9.464 ± 5.384, and 9.641 ± 5.382 respectively. T- test statistical analysis shows that there are no significant differences between the mean of canal transportation angles in two groups as measured by Schneider and Weine method (P>0.05). Mann- Whitney test shows that there are statistically significant differences between two groups as measured by LAT and Over Lay techniques (P<0.001). Conclusions: Results of this study shows that patency file in conjunction with P.S.B techniques causes significant reduction in apical transportation angle. Shortcomings of Schneider method in determination of canal curvature and specially assessment of apical transportation after instrumentation is mentioned in several studies. Our study shows that both Schneider and Weine techniques are not reliable methods for transportation assessment when compared with Image Over Lay technique. The result of LAT Shows close proximity with Over Lay technique and can be used for evaluation of apical transportation as a simple and reliable technique. Keywords: Apical transportation, Passive step back, Apical patency, Schneider technique, Weine technique, Long axis technique, OverLay technique

New salen-type manganese(III) Schiff base complexes derived from meso -1,2-diphenyl-1,2-ethylenediamine: In vitro anticancer activity, mechanism of action, and molecular docking studies

Four new manganese(III) Schiff base complexes (1-4) were synthesized and characterized. The complexes have general formula MnClLx in which L represents a Schiff base ligand derived from condensation of meso-1,2-diphenyl-1,2-ethylenediamine with salicylaldehyde or its 3-OMe-, 5-Br-, or 5-OMe-derivatives (x = 1-4, respectively). The crystal structure of MnClL1 (1) was characterized by X-ray crystallography. The in vitro anticancer activity of these complexes was evaluated by MTT and apoptosis assays against human breast (MCF-7) and liver (Hep G2) cancer cells. The complexes exhibited considerable antiproliferative activity against both cell lines (IC50 = 10.8-21.02 M) comparable to cis-platin, except 4 (MCF-7). The highest activity was found for 1 with IC50 values of 13.62 M (MCF-7) and 10.8 M (Hep G2). Flow cytometry experiments showed that 1 induced apoptosis on MCF-7 tumor cell line. Docking simulations using AUTODOCK were also carried out. The results showed that all complexes fitted into the minor groove region of DNA. © 2015 Taylor & Francis

Synthesis and anti-intracellular Copper overload evaluation of Nanoconjugated D-penicillamine –Dendrimer in Wilson’s model cells

Background and Objective: Wilson’s disease (WD) is caused by mutation to the cooer-transporting gene ATP7B. Chelation therapy is the main protochol of treatment for patients with Wilson’s disease. D-penicillamine is one of the well-known chelator agants which is used in WD treatment but it can not enter into the intracellular space.This study was done to evaluate the synthesis and anti-intracellular Copper overload evaluation of Nanoconjugated D-penicillamine –Dendrimer in Wilson’s model cells. Methods: In this descriptive-analytic study, initially 0.01 mm polyethylene glycol (PEG) and 0.0018 mm citric acid, Dendrimer was synthesized. After purification by dialysis bag and lyophilization, 10mg dendrimer was conjugated to 3.3mg D-penicillamine. Nanoconjugated D-penicillamine-dendrimer was injected on Wilson’s model cells. After incubation and centrifugation intracellular measurement of copper concentration and FTIR test were done. Results: Copper accumulation significantly reduced in the HepG2 WD cell by Nanoconjugated D-penicillamine - Dendrimer in compared to D-penicillamine (P<0.05). Copper accumulation was determined to be 46.61. MTT assay showed no toxicological damage in HepG2 WD cell. Conclusion: Nanoconjugated D-penicillamine –Dendrimer can reduces intracellular concentration of Copper

Evaluation of Platelet-Rich Plasma in Combination with Deproteinized Bovine Bone Mineral in the Rabbit Cranium; A Pilot Study

Statement of Problem: Reconstruction methods are an essential prerequisite for functional rehabilitation of the stomatognathic system. Platelet-rich plasma (PRP) offers a new and potentially useful adjunct to bone substitute materials (e.g. Xenografts) inoral and maxillofacial bone and implant reconstructive surgery.Purpose: An animal study was carried out to investigate the influence of PRP on the regeneration of non-critical sized bony defects, treated with Deproteinized Bovine Bone Mineral (DBBM).Materials and Methods: Eight New Zealand white rabbits were included in this randomized, pilot study. Three equal cranial bone defects (3×6 mm) were created and immediately grafted with DBBM and PRP+DBBM; one of them was left unfilled to serve as a control. The defects were evaluated using histologic and histomorphometricanalysis at 2, 4, 8 and 12 weeks.Results: The histomorphometric findings showed a significant increase in bone area and trabecular maturity in experimental defects as compared to the control at 4, 8 and 12 week intervals. A significant increase in bone formation was seen with the additionof PRP to DBBM at 2, 4 and 8 week intervals. At 12 weeks, the level of bone formation was similar between the two groups. There was also a significant increase in the rate of biodegradation of the DBBM particles with the addition of PRP at 2, 4, 8 and 12 weeks.No foreign body reaction and severe inflammation was seen in any of specimens.Conclusion: Within the limitations of this pilot study, it was concluded that the addition of PRP to Xenogenic bone substitute material in non-critical-sized defects of the rabbit cranium showed a histomorphometric increase in bone formation (until the 8th week ofhealing) and a greater amount of biomaterial degradation throughout the study period

Extending the application of a magnetic PEG three-part drug release device on a graphene substrate for the removal of Gram-positive and Gram-negative bacteria and cancerous and pathologic cells

M Ramezani Farani,1 P Khadive Parsi,1 Gh Riazi,2 M Shafiee Ardestani,3 H Saligeh Rad4,5 1School of Chemical Engineering, University College of Engineering, University of Tehran, Tehran 4563-11155, Iran; 2Institute of Biophysics and Biochemistry, University of Tehran, Tehran 1417614411, Iran; 3Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran; 4Quantitative Medical Imaging Systems Group, Research Center for Molecular and Cellular Imaging, Tehran University of Medical Sciences, Tehran, Iran; 5Medical Physics and Biomedical Engineering Department, Tehran University of Medical Sciences, Tehran, Iran Objective: In this study, novel graphene oxide (GO)-based nanocomposites are presented. In fact, we have tried to replace the carboxyl groups on the surface of GO with amine groups to allow the biocompatible poly(ethylene glycol) bis(carboxymethyl) ether (average Mn 600) polymer to bond through an amide bond. Materials and methods: The synthesis was conducted accurately according to final characterization experiments (Raman, X-ray diffraction [XRD], atomic force microscopy [AFM], X-ray photoelectron spectroscopy [XPS], thermogravimetric analysis [TGA], etc). The antimicrobial property of this nanocomposite was examined in Escherichia coli (ATCC 25922) as Gram-negative and Staphylococcus aureus (ATCC 25923) as Gram-positive bacterial species. Besides, curcumin (CUR) was added to the produced nanocomposite both as a promising anticancer drug and an antioxidant, the toxicity of which was then assessed on cellular-based HepG2 and pC12. Results: An intense increase in toxicity was detected by MTT assay. Conclusion: It can mainly be concluded that the nanocomposite synthesized in this study is capable of delivering drugs with antibacterial properties. Keywords: graphene oxide, magnetic nanocomposite, drug delivery, antimicrobial, curcumi

Mesoporous silica nanoparticles functionalized with folic acid/methionine for active targeted delivery of docetaxel

Pegah Khosravian,1 Mehdi Shafiee Ardestani,2 Mehdi Khoobi,3 Seyed Naser Ostad,4 Farid Abedin Dorkoosh,1 Hamid Akbari Javar,1,* Massoud Amanlou5,6,* 1Department of Pharmaceutics, 2Department of Radiopharmacy, 3Department of Pharmaceutical Biomaterials, 4Department of Pharmacology and Toxicology, 5Department of Medicinal Chemistry, Faculty of Pharmacy and Pharmaceutical Sciences Research Center, 6Drug Design and Development Research Center, Tehran University of Medical Sciences, Tehran, Iran *These authors contributed equally to this work Abstract: Mesoporous silica nanoparticles (MSNs) are known as carriers with high loading capacity and large functionalizable surface area for target-directed delivery. In this study, a series of docetaxel-loaded folic acid- or methionine-functionalized mesoporous silica nanoparticles (DTX/MSN-FA or DTX/MSN-Met) with large pores and amine groups at inner pore surface properties were prepared. The results showed that the MSNs were successfully synthesized, having good pay load and pH-sensitive drug release kinetics. The cellular investigation on MCF-7 cells showed better performance of cytotoxicity and cell apoptosis and an increase in cellular uptake of targeted nanoparticles. In vivo fluorescent imaging on healthy BALB/c mice proved that bare MSN-NH2 are mostly accumulated in the liver but MSN-FA or MSN-Met are more concentrated in the kidney. Importantly, ex vivo fluorescent images of tumor-induced BALB/c mice organs revealed the ability of MSN-FA to reach the tumor tissues. In conclusion, DTX/MSNs exhibited a good anticancer activity and enhanced the possibility of targeted drug delivery for breast cancer. Keywords: targeted delivery, mesoporous silica nanoparticle, folic acid, methionine, docetaxe

Crystal structures and in vitro anticancer studies on new unsymmetrical copper(II) Schiff base complexes derived from meso-1,2-diphenyl-1,2-ethylenediamine: a comparison with related symmetrical ones

Two new unsymmetrical copper(II) Schiff base complexes, CuLn(py)ClO4 (n = 1, 2) in which Ln represents a tridentate N2O type Schiff base ligand, were synthesized. Lns were derived from monocondensation of meso-1,2-diphenyl-1,2-ethylenediamine with salicylaldehyde or 3-methoxysalicylaldehyde. The reaction between CuLn(py)ClO4 and other salicylaldehyde derivatives resulted in new N2O2 unsymmetrical tetradentate CuII complexes, CuL3�6. Crystal structures of CuL1(py)ClO4, CuL4, and CuL5 were obtained. These new complexes as well as a series of related symmetrical ones (i.e. CuL7�12) were tested for their in vitro anticancer activity against human liver cancer cell line (Hep-G2) by MTT and apoptosis assay. All of the complexes showed considerable cytotoxic activity against tumor cell lines (IC50 = 5.13�16.24 μg mL�1). The symmetrical CuL7 was the most potent anticancer derivative (IC50 = 5.13 μg mL�1) compared to the control drug 5-FU (IC50 = 5.4 μg mL-1, p &amp;#x003C; 0.05). Flow cytometry experiments showed that the copper derivatives especially CuL2(py)ClO4 and CuL7 induced more apoptosis on Hep-G2 tumor cell lines compared to 5-FU. © 2016 Informa UK Limited, trading as Taylor & Francis Group