فهرست مطالب

Advanced Pharmaceutical Bulletin - Volume:7 Issue: 2, Jun 2017

Advanced Pharmaceutical Bulletin
Volume:7 Issue: 2, Jun 2017

  • تاریخ انتشار: 1396/03/01
  • تعداد عناوین: 20
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  • Sara Aqmasheh, Karim Shamsasanjan, Parvin Akbarzadehlaleh, Davod Pashoutan Sarvar, Hamze Timari Pages 165-177
    Hematopoiesis is a balance among quiescence, self-renewal, proliferation, and differentiation, which is believed to be firmly adjusted through interactions between hematopoietic stem and progenitor cells (HSPCs) with the microenvironment. This microenvironment is derived from a common progenitor of mesenchymal origin and its signals should be capable of regulating the cellular memory of transcriptional situation and lead to an exchange of stem cell genes expression. Mesenchymal stem cells (MSCs) have self-renewal and differentiation capacity into tissues of mesodermal origin, and these cells can support hematopoiesis through release various molecules that play a crucial role in migration, homing, self-renewal, proliferation, and differentiation of HSPCs. Studies on the effects of MSCs on HSPC differentiation can develop modern solutions in the treatment of patients with hematologic disorders for more effective Bone Marrow (BM) transplantation in the near future. However, considerable challenges remain on realization of how paracrine mechanisms of MSCs act on the target tissues, and how to design a therapeutic regimen with various paracrine factors in order to achieve optimal results for tissue conservation and regeneration. The aim of this review is to characterize and consider the related aspects of the ability of MSCs secretome in protection of hematopoiesis.
    Keywords: Hematopoietic Stem Cell, Cytokine, Mesenchymal Stem Cells, Microvesicle, miRNA
  • Roghiyeh Pashaei-Asl, Khodadad Khodadadi, Fatima Pashaei-Asl, Gholamreza Haqshenas, Nasser Ahmadian, Maryam Pashaiasl, Reza Hajihosseini Baghdadabadi Pages 179-187
    Finding novel and effective antibiotics for treatment of Legionella disease is a challenging field. Treatment with antibiotics usually cures Legionella infection; however, if the resultant disease is not timely recognized and treated properly, it leads to poor prognosis and high case fatality rate. Legionella pneumophila DrrA protein (Defects in Rab1 recruitment protein A)/also known as SidM affects host cell vesicular trafficking through modification of the activity of cellular small guanosine triphosphatase )GTPase( Rab (Ras-related in brain) function which facilitates intracellular bacterial replication within a supporter vacuole. Also, Legionella pneumophila LepA and LepB (Legionella effector protein A and B) proteins suppress host-cell Rab1 protein’s function resulting in the cell lysis and release of bacteria that subsequently infect neighbour cells. Legionella readily develops resistant to antibiotics and, therefore, new drugs with different modes of action and therapeutic strategic approaches are urgently required among antimicrobial drug therapies;gene therapy is a novel approach for Legionnaires disease treatment. On the contrary to the conventional treatment approaches that target bacterial proteins, new treatment interventions target DNA (Deoxyribonucleic acid), RNA (Ribonucleic acid) species, and different protein families or macromolecular complexes of these components. The above approaches can overcome the problems in therapy of Legionella infections caused by antibiotics resistance pathogens. Targeting Legionella genes involved in manipulating cellular vesicular trafficking using a dendrimer-mediated antisense therapy is a promising approach to inhibit bacterial replication within the target cells.
    Keywords: Legionella pneumophila, DrrA (SidM), LepA, LepB, Intracellular replication, Rab, Vesicular trafficking
  • Dariush Shanehbandi, Jafar Majidi, Tohid Kazemi *, Behzad Baradaran, Leili Aghebati-Maleki, Farzaneh Fathi, Jafar Ezzati Nazhad Dolatabadi Pages 189-194
    Purpose
    Surface plasmon resonance (SPR) sensing confers a real-time assessment of molecular interactions between biomolecules and their ligands. This approach is highly sensitive and reproducible and could be employed to confirm the successful binding of drugs to cell surface targets. The specific affinity of monoclonal antibodies (MAb) for their target antigens is being utilized for development of immuno-sensors and therapeutic agents. CD20 is a surface protein of B lymphocytes which has been widely employed for immunotargeting of B-cell related disorders. In the present study, binding ability of an anti-CD20 MAb to surface antigens of intact target cells was investigated by SPR technique.
    Methods
    Two distinct strategies were used for immobilization of the anti-CD20 MAb onto gold (Au) chips. MUA (11-mercaptoundecanoic acid) and Staphylococcus aureus protein A (SpA) were the two systems used for this purpose. A suspension of CD20-positive Raji cells was injected in the analyte phase and the resulting interactions were analyzed and compared to those of MOLT-4 cell line as CD20-negative control.
    Results
    Efficient binding of anti-CD20 MAb to the surface antigens of Raji cell line was confirmed by both immobilizing methods, whereas this MAb had not a noticeable affinity to the MOLT-4 cells.
    Conclusion
    According to the outcomes, the investigated MAb had acceptable affinity and specificity to the target antigens on the cell surface and could be utilized for immunodetection of CD20-positive intact cells by SPR method.
    Keywords: CD20, Surface Plasmon Resonance, Immobilization, Staphylococcus aureus protein A, 11-mercaptoundecanoic acid
  • Ebrahim Salehifar *, Shima Ebrahim, Mohammad-Reza Shiran, Fatemeh Faramarzi, Hossein Askari Rad, Razieh Avan, Asadollah Mohseni Kiasari, Pouneh Ebrahimi Pages 195-202
    Purpose
    Propranolol is the most widely used treatment for cardiovascular diseases. Dosage range in our patients is usually less than the amount mentioned in references. The aim of the present study was to clarify whether pharmacokinetic differences are able to justify the need for the fewer doses in our patients or not.
    Methods
    Twenty healthy volunteers (10 male) at heart center of Mazandaran University of Medical Sciences were studied. Samples of blood were collected before a single oral dose (40 mg) of Propranolol. Blood samples were taken up to 9 hours after dose. Total plasma concentration of Propranolol was measured by HPLC. Population Pharmacokinetic analysis was performed using population pharmacokinetics modeling software P-Pharm.
    Results
    The mean value for oral plasma clearance (CL/F) was 126.59 ml/hr. The corresponding values for apparent volume of distribution (V/F), t1/2 beta, maximum blood concentration (C max), and time to reach the maximum blood concentration (T max) were 334.12 Lit, 1.98 hr, 40.25 ng/ml, and 1.68 hr, respectively. The observed mean values of V/F of propranolol in the present study were comparable with those reported in the literature. However, the mean values of CL/F of propranolol in current study was significantly higher than those reported in other population (P-value
    Conclusion
    This study has confirmed that the pharmacokinetic differences are not able to justify over-responsiveness of Iranian population to propranolol. Pharmacodynamic differences in responding to beta blocker drugs by Renin secretion or having a different sensibility to beta receptors might play a role in making our population have a different response to propranolol.
    Keywords: Propranolol, Pharmacokinetics, Pharmacodynamic, Iranian Population, Polymorphism
  • Swati Jagdale*, Apoorva Chandekar Pages 203-213
    Purpose
    Inflammatory bowel disease (IBD) is a chronic, relapsing and often life-long disorder. The best way to tackle IBD is to develop a site targeted drug delivery. Methylprednisolone is a potent anti-inflammatory steroid. The relative potency of methylprednisolone to hydrocortisone is at least four is to one. The aim of the present research was to develop a colon targeted drug delivery for treatment of IBD.
    Methods
    Compression coated drug delivery system was designed and optimised. Core tablet contained drug, croscarmellose sodium (CCS-superdisintegrant), avicel (binder) and dicalcium phosphate (diluent). Design of experiment with 32 factorial design was applied for optimization of compression coated delivery. Chitosan and cellulose acetate phthalate were chosen as independent variables. Swelling index, hardness and % drug release were dependant variables.
    Results
    Core tablet (C5 batch) containing 2.15% CCS showed disintegration in less than 10sec. FTIR, UV and DSC study had shown absence of any significant physical and chemical interaction between drug and polymers. F8 was found to be optimised formulation. F8 contained 35% chitosan and 17.5% cellulose acetate phthalate. It showed drug release of 86.3% ± 6.1%, hardness 6.5 ± 1.5 and lag time 7 hrs. Simulated media drug release was 97.51 ± 8.6% with 7.5 hrs lag time. The results confirmed that the lag time was highly affected by the coating of the polymers as well as the concentration of the superdisintegrant used in core tablet.
    Conclusion
    In-vitro and in-vivo results confirmed a potential colon targeted drug therapy for treatment of IBD.
    Keywords: Methylprednisolone, Colon, Targeting, Press coated, Tablet, Chitosan
  • Sanaz Gholami, Seyede Momeneh Mohammadi, Ali Akbar Movasaghpour Akbari, Ali Abedelahi, Alireza Hemmati, Shirin Fallahi, Hojjatollah Nozad Charoudeh* Pages 215-220
    Purpose
    Terminal deoxynucleotidyl transferase(TdT) is a DNA polymerase that is present in immature pre-B and pre-T cells. TdT inserts N-nucleotides to the V (D) J gene segment during rearrangements of genes, therefore, it plays a vital role in the development and variation of the immune system in vertebrates. Here we evaluated the relationship between cytokines like interleukin-2 (IL-2), interleukin-7 (IL-7), and interleukin-15 (IL-15) and TdT expression in cord blood mononuclear cells and also effect of inhibition in the expansion of B and T cells derived from cord blood.
    Methodes: The cord blood mononuclear cells were cultured with different combination of cytokines for 21days, which they were harvested in definite days (7, 14 and 21) and evaluated by flow cytometry.
    Results
    Our data indicated that TdT expression increased in cord blood mononuclear cells using immune cell key cytokines without being dependent on the type of cytokines. TdT inhibition reduced both the expansion of B and T cells derived from cord blood and also declined the apoptosis and proliferation. Considered together, TdT played an important role in the control of the expansion of B and T cells derived from cord blood.
    Conclusion
    considered together, it was observed that TdT expression was increased by cytokines and TdT inhibition not only reduced B and Tcells derived from cord blood, but it also affected the rate of apoptosis and proliferation.
    Keywords: Cord blood, TdT inhibition, B, T cells, Cytokines, Genistin
  • Nazli Namazi, Mohammad Alizadeh, Elham Mirtaheri, Safar Farajnia Pages 221-228
    Purpose
    Obesity is a multi-factorial health problem which results from the interaction of environmental and genetic factors. The aim of the present study was to determine the effects of dried licorice extract with a calorie restricted diet on anthropometric indices and insulin resistance with nutrigenetic approach.
    Methods
    For this pilot, double-blind, placebo-controlled randomized clinical trial, 72 eligible subjects were randomly allocated to Licorice or placebo group. They received a low-calorie diet either with a 1.5 g/day of Licorice extract or placebo for 8 weeks.
    Results
    There were no significant differences in anthropometric indices and dietary intake in genotype subgroups at the baseline. Findings indicated that supplementation with Licorice extract did not change anthropometric indices and biochemical parameters significantly compared to a hypocaloric diet alone. However, from the nutrigenetic point of view, significant changes in anthropometric indices and QUICKI were observed in the Pro12Pro genotypes compared to the Pro12Ala at the end of the study (p
    Conclusion
    In obese subjects, the Pro/Pro polymorphism of the PPAR-γ2 gene seems to induce favourable effects on obesity management. Further studies are needed to clarify whether PPAR-γ2 gene polymorphisms or other obesity genes can affect responses to obesity treatment.
    Keywords: Licorice, Hypocaloric diet, Nutrigenetics, PPAR-γ2
  • Swati Jagdale *, Saylee Pawar Pages 229-239
    Purpose
    Ofloxacin is a fluoroquinolone with broad-spectrum antibacterial action, used in treatment of systemic and local infections. Ofloxacin is BCS class II drug having low solubility, high permeability with short half-life. The present work was aimed to design, develop and optimize gellified emulsion of Ofloxacin to provide site targeted drug delivery. Transdermal drug delivery will enhance the bioavailability of the drug giving controlled drug release.
    Methods
    Transdermal drug delivery system was designed with gelling agent (Carbopol 940 and HPMC K100M), oil phase (oleic acid) and emulsifying agent (Tween 80: Span 80). Effect of concentration of gelling agent on release of drug from transdermal delivery was studied by 32 factorial design. Emulgel was evaluated for physical appearance, pH, drug content, viscosity, spreadability, antimicrobial activity, in- vitro diffusion study and ex-vivo diffusion study.
    Results
    FE-SEM study of the emulsion batch B5 has revealed formation of emulsion globules of approximately size 6-8 µm with -11.2 mV zeta potential showing good stability for the emulsion. Carbopol 940 had shown greater linear effect on drug release and viscosity of the formulations due to its high degree of gelling. In-vitro diffusion study through egg membrane had shown 88.58±1.82 % drug release for optimized batch F4. Exvivo diffusion study through goat skin indicated 76.68 ± 2.52% drug release.
    Conclusion
    Controlled release Ofloxacin emulgel exhibiting good in-vitro and ex-vivo drug release proving good antimicrobial property was formulated.
    Keywords: Emulgel, Transdermal, Delivery, Ofloxacin, Antimicrobial, Emulsion
  • Roghiyeh Pashaei-Asl, Fatima Pashaei-Asl, Parvin Mostafa Gharabaghi, Khodadad Khodadadi, Mansour Ebrahimi, Esmaeil Ebrahimie, Maryam Pashaiasl * Pages 241-249
    Purpose
    Ginger is a natural compound with anti-cancer properties. The effects of ginger and its mechanism on ovarian cancer and its cell line model, SKOV-3, are unclear. In this study, we have evaluated the effect of ginger extract on SKOV-3.
    Methods
    SKOV-3 cells were incubated with ginger extract for 24, 48 and 72 hours. Cell toxicity assay was performed. Different data mining algorithms were applied to highlight the most important features contributing to ginger inhibition on the SKOV-3 cell proliferation. Moreover, Real-Time PCR was performed to assay p53, p21 and bcl-2 genes expression. For co-expression meta-analysis of p53, mutual ranking (MR) index and transformation to Z-values (Z distribution) were applied on available transcriptome data in NCBI GEO data repository.
    Results
    The ginger extract significantly inhibited cancer growth in ovarian cancer cell line. The most important attribute was 60 µg/ml concentration which received weights higher than 0.50, 0.75 and 0.95 by 90%, 80% and 50% of feature selection models, respectively. The expression level of p53 was increased sharply in response to ginger treatment. Systems biology analysis and meta-analysis of deposited expression value in NCBI based on rank of correlation and Z-transformation approach unraveled the key co-expressed genes and coexpressed network of P53, as the key transcription factor induced by ginger extract. High co-expression between P53 and the other apoptosis-inducing proteins such as CASP2 and DEDD was noticeable, suggesting the molecular mechanism underpinning of ginger action.
    Conclusion
    We found that the ginger extract has anticancer properties through p53 pathway to induce apoptosis.
    Keywords: Ovarian cancer, Ginger extract, Anticancer, P53, Bcl-2, Systems biology analysis, Meta-analysis
  • Cristina Mateescu, Anca Mihaela Popescu*, Gabriel Lucian Radu, Tatiana Onisei, Adina Elena Raducanu Pages 251-259
    Purpose
    This study was carried out in order to find a reliable method for the fast detection of adulterated herbal food supplements with sexual enhancement claims. As some herbal products are advertised as “all natural”, their “efficiency” is often increased by addition of active pharmaceutical ingredients such as PDE-5 inhibitors, which can be a real health threat for the consumer.
    Methodes: Adulterants, potentially present in 50 herbal food supplements with sexual improvement claims, were detected using 2 spectroscopic methods - Raman and Fourier Transform Infrared - known for reliability, reproductibility, and an easy sample preparation. GC-MS technique was used to confirm the potential adulterants spectra.
    Results
    About 22% (11 out of 50 samples) of herbal food supplements with sexual enhancement claims analyzed by spectroscopic and spectrometric methods proved to be "enriched" with active pharmaceutical compounds such as: sildenafil and two of its analogues, tadalafil and phenolphthalein. The occurence of phenolphthalein could be the reason for the non-relevant results obtained by FTIR method in some samples. 91% of the adulterated herbal food supplements were originating from China.
    Conclusion
    The results of this screening highlighted the necessity for an accurate analysis of all alleged herbal aphrodisiacs on the Romanian market. This is a first such a screening analysis carried out on herbal food supplements with sexual enhancement claims.
    Keywords: Fourier transform infrared, GC-MS, Herbal food supplements, PDE-5 inhibitors, Raman spectroscopy
  • Parizad Piran, Hossein Samadi Kafil, Saeed Ghanbarzadeh, Rezvan Safdari, Hamed Hamishehkar * Pages 261-268
    Purpose
    Due to the antimicrobial property, menthol have significant potential for food preservation and foodstuff shelf life improvement. Nevertheless, menthol instability, insolubility, and rapid crystallization in aqueous media make it unsuitable for used in food products. This work was aimed to prepare menthol-loaded nanostructured lipid carriers (NLCs) to enhance its antimicrobial activity.
    Methods
    Morphology, particle size and size distribution, encapsulation efficiency percent (EE%), and physical stability of the optimized formulation, prepared by hot melt homogenization method, were characterized by scanning electron microscopy, particle size analyzing, gas chromatography, and X-ray diffraction (XRD) methods. Minimum inhibitory concentration and minimum bactericidal concentration of menthol-loaded NLCs were evaluated and compared with conventional menthol emulsion against various Gram-positive (Staphylococcus aureus, Bacillus cereus) and Gram-negative bacteria (Escherichia coli), as well as one fungus (Candida albicans).
    Results
    Menthol-loaded NLCs were spherically shaped nanosized (115.6 nm) particles with narrow size distribution (PDI = 0.2), suitable menthol EE% (98.73%), and appropriate physical stability after 90 days of storage period. XRD results indicated that menthol was in the amorphous form in the nanoparticles matrix. Antibacterial assay results revealed that the menthol-loaded NLCs exhibited significantly higher in vitro antimicrobial property than conventional menthol emulsion. The results also indicated that menthol-loaded NLCs had better effect on fungi than bacteria, and furthermore, antibacterial efficiency on Grampositive bacteria was higher than Gram-negative bacteria.
    Conclusion
    In conclusion, NLCs could be a promising carrier for improvement of antimicrobial activity and preservation efficacy of essential oils in foodstuffs.
    Keywords: Menthol, Nanostructure lipid carriers, NLC, Antimicrobial activity, Food preservative
  • Farideh Doostan, Roxana Vafafar, Parvin Zakeri-Milani, Aliasghar Pouri, Rogayeh Amini Afshar, Mehran Mesgari Abbasi * Pages 269-274
    Purpose
    Methotrexate (MTX) is prescribed in many diseases and can result in oxidative stress (OS) followed by injuries in some tissues. Antioxidants administration are effective in reducing OS. Pomegranate exhibits high anti-oxidant capacities. This study investigated whether pomegranate seed and peel methanolic extracts (PSE and PPE) could protect against MTX-induced OS and lipid profile changes in rats.
    Methods
    Forty-eight rats were randomly divided into 6 groups: control group (normal salin), PSE group (500 mg/kg, orally), PPE group (500 mg/kg, orally), MTX group (10 mg/kg, IM), MTX and PSE group, and MTX and PPE group. Blood samples were taken for analysis in the end of the procedure.
    Results
    The findings showed a significant reduction in Glutathione peroxidase (GPx) and Superoxide dismutase (SOD), and an enhancement in malondialdehyde (MDA) values after MTX treatment (p
    Conclusion
    The results showed the OS induced by MTX and the protective effects of PSE and PPE against MTX-induced serum oxidative stress and lipid profile changes in rats.
    Keywords: Methotrexate, Oxidative stress, Pomegranate, Rats
  • Alireza Farnam, Arezoo Mehrara*, Hossein Dadashzadeh, Golamreza Chalabianlou, Salman Safikhanlou Pages 275-279
    Purpose
    Depression causes dysfunction in various spheres of individual and social life, which is now considered as the fourth-leading cause of the global disease burden. Given that violence and aggression associated with depression in the community cause serious damage to the family, the prediction, early detection and effective treatment of aggressive and violent behavior are essential. The present study compared the severity of aggression before and after treatment with sertraline in patients with major depression.
    Methods
    This is an intervention type study and the study population consisted of patients with depression and aggression. The sampling included 23 eligible patients. Data were obtained by SCID-I, SCID-II, STAXI-II, BDI-II and was also analyzed using SPSS 23 software.
    Results
    The results showed that depression, anger mood, desire to verbally express anger, controlling anger and anger control before treatment was reduced but the desire for physical expression of anger increased.
    Conclusion
    Obtained results in this research support the effect of Sertraline on reduction of severity of depression, reduction of severity of symptoms of aggression and anger (state of anger, anger feeling, and the tendency to express anger verbally), increased controlling external anger and significantly controlling internal anger. Hence, Sertraline can be found effective in the treatment of patients with depression and aggressive behaviors. Also Sertraline increases tend to cause physical representation of anger, then this issue supports the increase in the euthanasia behavior in primary days of treatment with SSRI that requires more assessments.
    Keywords: Depression, Aggression, Sertraline, STAXI-II
  • Eskandar Moghimipour, Anayatollah Salimi *, Sahar Changizi Pages 281-289
    Purpose
    The objective of the present study is to formulate and evaluate a new microemulsion (ME) for topical delivery of griseofulvin.
    Methods
    The solubilities of griseofulvin in different combinations of surfactant to cosurfactant (S/Co ratio) were determined. Accordingly, based on their phase diagrams, eight microemulsions were formulated and then evaluated with respect to their particle size, surface tension, viscosity, conductivity, zeta potential and stability. Their release behavior, Scanning Electron Microscopy (SEM), Differential Scanning Calorimetry (DSC), refractory index (RI), pH and Small-angle-X-ray scattering (SAXS) were also assessed.
    Results
    The results indicated that the mean droplet size of the MEs ranged from 30.9 to 84.3 nm. Their zeta potential varied from -4.5 to -20.8. Other determined characteristics were viscosity: 254-381 cps, pH: 5.34-6.57, surface tension: 41.16- 42.83 dyne.cm-1, conductivity: 0.0442 – 0.111 ms.cm-1. The drug release was in the range of 22.4 to 43.69 percent. Also, hexagonal, cubic and lamellar liquid crystals were observed in SAXS experiments.
    Conclusion
    It can be concluded that any alteration in MEs constituents directly affects their microstructure, shape, droplet size and their other physicochemical properties.
    Keywords: Griseofulvin, Microemulsion, Microstructure, XRD, DSC, Stability
  • Nastaran Faghihi, Mohammad Taghi Mohammadi * Pages 291-298
    Purpose
    The pleiotropic effects of statins (antioxidant and anti-inflammation) have been reported by previous studies. Therefore, we aimed to determine whether pitavastatin has protective effects against pentylenetetrazol (PTZ)-induced kindling in mice and also whether pitavastatin improves the brain antioxidant capacity and attenuates the oxidative injuries in kindled mice.
    Methods
    Twenty-four mice were randomly divided into four groups (each group n=6); control, PTZ-kindling and PTZ-kindled rats treated with pitavastatin (1&4 mg/kg). PTZ kindling seizures were induced by repetitive intraperitoneal injections of PTZ (65 mg/kg) every 48 hours till day twenty-one. Animals received daily oral pitavastatin for twenty-one days. Latency, score and duration of the seizures were recorded. The activities of catalase (CAT) ad superoxide dismutase (SOD), and likewise the contents of malondialdehyde (MDA) and nitrate were assessed in the brains of all rats.
    Results
    There was a progressive reduction in latency of the kindled rats in the next injections of PTZ. Pitavastatin reduced this value (latency) particularly at higher dose. Seizures duration and score also decreased in treatment groups. SOD and CAT activities significantly decreased in PTZ-kindling group by 62% and 64%, respectively, but pitavastatin did not significantly change the SOD and CAT activities. Brain MDA and nitrate significantly increased in PTZ-kindling group by 53% and 30%, respectively. Pitavastatin at higher dose significantly decreased the MDA and nitrate contents of PTZkindling rats by 45% and 32%, respectively.
    Conclusion
    Our findings revealed that pitavastatin can improve the behavioral expression of the PTZ-kindling rats and attenuate the seizure-induced oxidative/nitrosative damage.
    Keywords: Seizure, Oxidative damage, Nitrosative damage, Pentylenetetrazole, Pitavastati
  • Mohammad Tasyriq Che Omar * Pages 299-312
    Purpose
    More than half of the diagnostic and therapeutic recombinant protein production depends on mammalian-based expression system. However, the generation of recombinant antibodies remains a challenge in mammalian cells due to the disulfide bond formation and reducing cytoplasm. Therefore, the production of functional recombinant antibodies in target cell line is necessary to be evaluated before used in therapeutic application such intrabodies against HIV-1.
    Methods
    The work was to test expression of a single-chain variable fragment (scFv) antibody against HIV-1 Capsid p24 protein in a human mammalian-based expression system using HEK293T and Jurkat T cells as a model. Three expression plasmid vectors expressing scFv 183-H12-5C were generated and introduced into HEK293T. Expression of the scFv was analyzed, while ELISA and immunoblotting analysis verified its binding. The evaluation of the recombinant antibody was confirmed by HIV-1 replication and MAGI infectivity assay in Jurkat T cells.
    Results
    Three plasmid vectors expressing scFv 183-H12-5C was successfully engineered in this study. Recombinant antibodies scFv (~29 kDa) and scFv-Fc (~52 kDa) in the cytoplasm of HEK293T were effectively obtained by transfected the cells with engineered pCDNA3.3-mu-IgGk-scFv 183-H12-5C and pCMX2.5-scFv 183-H12-5C-hIgG1-Fc plasmid vectors respectively. scFv and scFv-Fc are specifically bound recombinant p24, and HIV-1 derived p24 (gag) evaluated by ELISA and Western blot. Jurkat T cells transfected by pCDNA3.3-scFv 183-H12-5C inhibit the replication-competent NL4-3 viral infectivity up to 60%.
    Conclusion
    Anti-p24 scFv 183-H12-5C antibody generated is suitable to be acted as intrabodies and may serve as a valuable tool for the development of antibody-based biotherapeutics against HIV-1.
    Keywords: Single-chain variable fragment (scFv), scFv-Fc, HIV-1 Capsid p24 protein, Mammalian-based expression, Intrabody
  • Qais Ibrahim Abualassal *, Khaldun Mohammad Al Azzam, Ziad Helmi Abudayeh, Loay Khaled Hassouneh Pages 313-321
    Purpose
    To describe a chemoenzymatic approach joining an enzymatic regioselective hydrolysis of peracetylated N-acetyl-α-D-glucosamine (A) with a mild controlled acyl relocation which resulted 2-acetamido-2 deoxy-1,3,6-tri-O-acetyl-α-D-glucopyranose (1B).
    Methods
    Immobilization of lipase on decaoctyl (DSEOD) and octyl-agarose (OSCL) was carried out as reported by the work of Bastida et al. The newly developed RP-HPLC method for examining the enzymatic hydrolysis was carried out isocratically utilizing a HPLC system.
    Results
    The new approach resulted the target compound (B) in 95% yield after purification utilizing flash column chromatography. Candida rugosa-lipase immobilized ondecaoctylsepabeads was the best catalyst in terms of activity and region-selectivity in the hydrolysis of substrate (A), delivering the deacetylation at C6 position (98% general yield). Also, a reversed-phase high-performance liquid-chromatographic (RP-HPLC) method for controlling the region-selective hydrolysis of peracetylated N-acetyl-α-D-glucosamine (A) with a mild monitored acyl movement which led to 2-acetamido-2-deoxy-1,3,6-tri-O-acetyl-αD-glucopyranose (1B) has additionally been developed. The developed RP-HPLC method was utilized as fingerprints to follow the hydrolysis of substrate (A) and to determine its purity and additionally yield. Furthermore, the acquired compound (B) was further purified by flash chromatography. Compound (B) was further characterized utilizing 1HNMR and mass spectrometry.
    Conclusion
    An efficient chemoenzymatic procedure to optimize the preparation of peracetylated lactosamine Bcontaining acetyl ester as extraordinary protecting group is presented. Compound Bis a significant intermediate for the synthesis of pharmacologically active compound (e.g. complex oligosaccharides for biochemical, biophysical, or biological examinations). Besides, reaction monitoring utilizing HPLC proposes more exact information than spectroscopic methods.
    Keywords: Glycosylation reaction, Chemo-enzymatic synthesis, Regioselectivity, High performance liquid chromatography, Flash column chromatography, Candida rugosa
  • Mehrnaz Tabibian, Mohammadali Torbati, Mohammad Reza Afshar Mogaddam, Maryam Mirlohi, Malihe Sadeghi, Javad Mohtadinia * Pages 323-328
    Purpose
    Vitamin D, a fat-soluble secosteroid, has a significant role in bone metabolism and helps calcium absorption in the body. Since vitamin D concentration is altered in fortified foods and dietary supplements, the actual amount of vitamin D may differ from the label value.
    Methods
    In this study, the concentrations of vitamin D2 and D3 of fortified bread sample were analytically determined. For this purpose, dough or homogenized bread sample was saponified using potassium hydroxide solution (30%, w/v) at 80°C, and the saponified analytes were extracted into n-heptane followed by liquid-liquid extraction. Then n-heptane fraction was evaporated to dryness and the sample was reconstituted in methanol. The effect of different parameters was evaluated by one variable at one-time strategy.
    Results
    The analytes concentrations were evaluated in dough fermentation, baking and storage steps. The effect of temperature in dough fermentation and baking was evaluated at the range of 5-30 and 200-250°C, respectively. Also, the fermentation time was studied in the range of 0-120 min. The analytes concentrations were followed for 1 to 5 days after baking. The results indicated that dough fermentation temperature has no significant effect on the concentration of the analytes. On the other hand, when the dough fermentation time and baking temperature are increased, the analytes concentrations are decreased. Also, the storage duration of the spiked bread samples decreased the analytes concentrations after one day.
    Conclusion
    Based on the obtained results, baking the dough at high temperatures lead to decrease in vitamin levels.
    Keywords: Bread sample, Vitamin D3, Vitamin D2, High performance liquid chromatography
  • Ali Kamal Attia *, Ahmed Sayed Saad, Manal Sami Alaraki, Eman Saad Elzanfaly Pages 329-334
    Purpose
    Thermal analysis techniques have been applied to study the thermal behavior of fenbendazole (Fen) and rafoxanide (Raf). Semi-empirical molecular orbital calculations were used to confirm these results.
    Methods
    Thermogravimetric analysis, derivative thermogravimetry, differential thermal analysis and differential scanning calorimetry were used to determine the thermal behavior and purity of the drugs under investigation.
    Results
    Thermal behavior of Fen and Raf were augmented using semi-empirical molecular orbital calculations. The purity values were found to be 99.17% and 99.60% for Fen and Raf, respectively.
    Conclusion
    Thermal analysis techniques gave satisfactory results to obtain quality control parameters such as melting point and degree of purity at low cost, furthermore, its simplicity and sensitivity justifies its application in quality control laboratories.
    Keywords: Fenbendazole, Rafoxanide, Thermal analysis, Purity, Molecular orbital calculations
  • Iwan Budiwan Anwar *, Asep Santoso, Eko Saputra, Rifky Ismail, J. Jamari, Emile Van Der Heide Pages 335-338
    Purpose
    Human bone marrow-derived mesenchymal cell (hBMC) reactions to 316L stainless steel (316L-SS) have never been evaluated. The objective of this study was to assess cell viability and interleukin-6 expression of hBMC cultures upon treatment with a 316L-SS implant.
    Methods
    A cytotoxicity analysis was conducted with a 3-(4,5-dimethylthiazol 2-yl)-2,5diphenyltetrazolium (MTT) assay after a period of 24, 48 and 72 hours of incubation. Expression of interleukin-6 was measured using enzyme-linked immunosorbent assay (ELISA).
    Results
    Cell viability measurement was performed via IC50 formula. All treatment group showed a > 50 % cell viability with a range of 56,5 - 96,9 % at 24 hours, 51,8-77,3% at 48 hours and 70,1- 120 % at 72 hours. Interleukin-6 expression was downregulated subsequent to treatment with 316L-SS compared to the control group.
    Conclusion
    We found that 316L-SS did not exhibit toxicity towards hBMC culture.
    Keywords: 316L stainless steel, Human bone marrow-derived mesenchymal cells, Cell viability, Interleukin-6