فهرست مطالب

Applied Biotechnology Reports - Volume:8 Issue: 1, Winter 2021
  • Volume:8 Issue: 1, Winter 2021
  • تاریخ انتشار: 1400/02/05
  • تعداد عناوین: 12
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  • Fariba Zafari, Morteza Sadeghi * Pages 1-4
    Introduction

    Matrix metalloproteinases (MMPs) are a family of zinc-dependent proteinase. Several studies have reported the association between the single nucleotide polymorphisms(C/T) -1562 in the MMP-9 promoter and the risk of cancer. In this study, we decided to carry out a comprehensive meta-analysis to obtain a reasonable result about the association between this polymorphism and the risk of lung cancer.

    Materials and Methods

    A complete literature review was conducted within the databases of ISI Web of Knowledge, google scholar, and PubMed for studies on lung cancer published from 2002 to 2018. A meta-analysis was conducted which included more than 3000 case and control subjects. The pooled Odds Ratio (OR) and 95% Confidence Intervals ( CI) were used for dominant, recessive, and co-dominant MMP-9 genotypes to assess the strength of the association.

    Results

    Analysis indicated no significant association between MMP-9-1562C/T polymorphism and the risk of lung cancer, dominant model; [CT+TT/CC]: OR = 0.972, 95% CI = 0.811–1.164, recessive model [CC+CT/TT] OR= 1.027, 95% CI = 0.651–1.618 and co-dominant model [TT/CC] OR = 0.983, 95% CI = 0.550–1.755.

    Conclusions

    No association was observed between the MMP9 -1562 C/T polymorphism and the incidence of lung cancer. The meta-analysis demonstrated that this polymorphism can't serve as a diagnostic marker for lung cancer.

    Keywords: MMP-9, Lung cancer, Polymorphism
  • Shima Jahanfar, Mehrdad Ghavami, Kianoush Khosravi Darani *, Mahshid Jahadi Pages 5-12
    Introduction

    Although Green tea is a rich source of antioxidant, use of this herbal in food industries is limited due to its oxygen and light instabilities.

    Materials and Methods

    Green tea polyphenols were encapsulated into liposomes using Mozafari method (with no solvents and detergents) to improve bioavailability of tea polyphenols. Screening design was used to find the major variables within all possible process variables. Then, optimal conditions were studied using response surface.

    Results

    The most appropriate condition was achieved using phosphatidylcholine of 4.5% (w/w), extract concentration of 0.7%, mixing time of 30 min and temperature of 50 °C. Encapsulation efficiency (EE) depended on concentrations of the green tea extract and phosphatidylcholine. The EE in optimal formulation was reached as 51.34%. The particle size and Z-potential of liposomal green tea extract were assessed at 419 nm and -59.7 mV, respectively. The total polyphenol content (TPC) of green tea included 164.2 mg gallic acid/g extract. Free radical scavenging activities of free and liposomal extracts were calculated as 90.6 and 93.37%, respectively, using 2-2-diphenyl-1-pycrylhydrazyl (DPPH) method.

    Conclusions

    Results revealed that liposomal green tea extract can be used extensively in food industries due to its high antioxidant activity. No size decreasing methods (e.g. sonication and homogenization) were needed to produce nanosize liposomal extracts to avoid structure instability.

    Keywords: Green Tea Extract, Liposome, polyphenols, Process variables, Plackett-Burmann Design, response surface method
  • Nour El Houda Bekkar *, Boumediene Meddah, Bahadir Keskin, Pascal Sonnet Pages 13-26
    Introduction

    The aim of this study was to evaluate the chemical composition of Z. lotus (ZL) and R. chalepensis (RC) Essential Oils (EOs), the oral acute toxicity, influence on the gastrointestinal (GI) microbiota, and the in vivo anti-salmonellosis effect.

    Materials and Methods

    The EOs were isolated using the steam distillation process, and bioactive components were identified by GC-MS analysis. Oral acute toxicity, influence on the GI flora composition, and the anti-salmonellosis effect were elucidated using in vivo methods on experimental animals.

    Results

    The GC-MS allowed us to identify 33 and 58 components in Z. lotus and R. chalepensis, respectively. Di-isooctyl phthalate (89.857%) was found to be the major compound identified in ZL. The main compounds in RC were 2-undecanone (26.528 %) followed by 2-nonanone (13.404 %). The LD50 of EOs was found to be greater than 5000 mg/kg. Also, no negative influence to intestinal microbiota was detected. An important decrease in S. enterica ssp arizonae cells achieving a bactericidal effect was recorded in rats treated with the EOs of both plants at a dose of 400 mg/kg. In parallel, an important significant (p <0.05) increase in lymphocyte number was observed for all tested animals. A decrease in alkaline phosphatase (ALP), alanine aminotransferase(ALT), and aspartate aminotransferase (AST) levels were observed. Furthermore, a reduced blood sedimentation rate (ESR) was recorded in treated animals.

    Conclusions

    The Z. lotus and R. chalepensis act effectively as anti-salmonellosis agents, which support the use of these plants to cure gastrointestinal infections.

    Keywords: Zizyphus lotus, Ruta chalepensis, Essential oils, Acute toxicity, Gastrointestinal microbiota, Anti-salmonellosis
  • Nedda Hamidi, Bahram Golestani Eimani, MohammadJavad Motamedi, Rouhollah Kazemi, Abbas Hajizade, Jafar Amani * Pages 27-31
    Introduction

    Enterotoxigenic E. coli (ETEC) is one of the major causes of watery diarrhea outbreaks in children under five years of age and passengers in developing countries. The pathogenicity of ETEC is due to the secretion of Colonization factors (Cfs) and two enterotoxins including heat-labile (Lt) and heat-stable (St). Although diarrhea is considered as one of the causes of mortality in developing countries, no approved vaccine is available for the disease caused by ETEC. Accordingly, the objective of the present study was to design a vaccine candidate containing Sta toxin which accounts for 30-75% of ETEC species.

    Materials and Methods

    A chimeric construct consisting of two Sta toxoid connected together by two linkers was designed. After expression and purification by Ni-NTA column, western blotting was performed to confirm the protein. The 2Sta protein was administered to BALB/c mice via injection and the serum and fecal antibodies titer was evaluated by the ELISA test.

    Results

    The recombinant 2Sta protein was expressed in an insoluble form (inclusion body) and the 20 kDa band was observed on the SDS-PAGE 12%. The results of the ELISA test suggested that IgG and IgA antibodies had enhanced compared to the control group.

    Conclusions

    The Sta protein which was produced in most ETEC species can be induced in the immune system and raised the serum and fecal antibodies. Using this candidate subunit vaccine could actually protect bacterial infection.

    Keywords: Heat stable (St) enterotoxin, Enterotoxigenic E. coli (ETEC), Diarrhea, Sta subunit, Vaccine
  • Maryam Mobarakian, Mehdi Nikbakht Dastjerdi, Jahanshir Shakarami *, Seyed Mohammad Abtahi Pages 32-36
    Introduction

    To the best of our knowledge, there is little information in regards to the cytotoxicity of 1,8- cineole. Insect cells exhibit wide resistance to the lethal effects of chemical compounds. Activation of p53 by acetylation can cause cell toxicity. This study has been carried out in order to investigate whether the use of 1,8-cineole in the Spodoptera frugiperda (SF-9) cell line can induce cytotoxicity by increasing p53 acetylation expression or not.

    Materials and Methods

    SF-9 cell line was cultured in TC100 insect culture medium and treated with 1,8-cineole at concentration of 850.45 μmol/L, based on the half-maximal inhibitory concentration (IC50) index at different times (24, 48, and 72h). The IC50 value was estimated for 1,8-cineole in SF-9. The percentage of alive cells was measured by MTT assay. The ELISA and Bradford protein techniques were used to detect endogenous levels of total and acetylated p53 protein generated in SF-9 cells.

    Results

    The findings of the present study indicated that treatment with 850.45 μmol per liter of 1,8-cineole shows a time-dependent increase in the number of dead cells of SF-9 cell line. The effect of severe toxicity on SF-9 was observed after 72 h of incubation with 1,8-cineole, with approximately 4% of SF-9 cells alive. We observed a significant increase in the level of p53 acetylation up to 48 h in SF-9, indicating that 1,8-cineole resulted in up‑regulation of acetylated P53 and consequently p53 activation in SF-9 cells. Results showed that there is relationship between acetylated p53 protein levels and 1,8-cineole toxicity in SF-9 cell line.

    Conclusions

    1,8-cineole, may function through common pathways and mediate their cytotoxic effects through targeting p53 and its acetylation in SF-9 cells.

    Keywords: 1, 8-cineole, p53, SF-9 Cell Line, Cytotoxicity
  • Akihito Nakanishi *, Yuri Sakihma, Nanami Ozawa Pages 37-40
    Introduction

     Chlamydomonas reinhardtii produces lipid and carbohydrate as an industrially useful bioproduct with the supply of CO2 as a carbon source. The CO2 supplying system, especially aeration rate through the photobioreactor, should be controlled to enhance cell proliferation.

    Materials and Methods

     After fixing CO2 concentration as 0.8%, the aeration rate was controlled to increase stepwisely by 10 mL·min-1, 20 mL·min-1, or 40 mL·min-1 beginning at 10 mL·min-1 to a maximum of 50 mL·min-1 after the pH ˃ 6.5. To show the effect of CO2-supply, the broth condition and the cell-component of lipid, carbohydrate, and protein were evaluated.

    Results

     The CO2 supplying condition increasing by 10 mL·min-1 stepwisely when over pH 6.5 in 100 mL of broth led to rapid cell proliferation reached a plateau 2 days earlier than in other conditions. On the other hand, the cell components incubated under 10 mL·min-1 stepwise condition showed no difference among the other conditions.

    Conclusions

     Cell proliferation was improved by optimized stepwise CO2 aeration rates versus cell concentration in broth, and cell components were not changed even with improved cell proliferation. According to the results, it could be possible to improve material productivity by increasing biomass productivity.

    Keywords: Chlamydomonas reinhardtii, Cell proliferation, CO2 supply
  • Nafiseh Ghorbani, Mehdi Assmar *, Nour Amirmozafari, Khosrow Issazadeh Pages 41-45
    Introduction

     Globally, Salmonella enterica serotype Typhi is responsible for more than 10 million enteric fever cases, annually. Because of the emergence of multidrug-resistant strains of many bacteria, including Salmonella Typhi, vaccination may be a preferred strategy to combat infectious diseases. In the present study, the efficiency of flagellin protein as a recombinant vaccine candidate was evaluated in BALB/c mice.

    Materials and Methods

    For this aim, flagellin protein was expressed in E. coli BL21 (DE3). Mice were grouped into two groups: test and control. The test group was immunized by the intraperitoneal administration of recombinant protein in combination with Freund's adjuvant. Following the completion of the immunization period, the mice were challenged by IP injection of 10 LD50 of live Salmonella Typhi and subsequent culture of their spleens and livers.

    Results

    Flagellin protein expression was confirmed by SDS-PAGE and Western blotting. ELISA showed the proper stimulation of the humoral immunity of the immunized mice. The bacterial count decreased significantly in the spleens and livers of the immunized animals in comparison to the control ones.

    Conclusions

    Findings of this study show the efficiency of flagellin recombinant protein in protecting mice against Salmonella Typhi.

    Keywords: Salmonella Typhi, Flagellin recombinant protein, Vaccine candidate, Enteric Fever
  • Mojtaba Sharti, Hadi Esmaeili Gouvarchin Ghaleh, Ruhollah Dorostkar *, Bahman Jalali Kondori Pages 46-50
    Introduction

    Antiviral therapy is an alternative for viral infection control when the virus is identified. As antiviral therapy has effectively used basic science to create very efficient treatments for severe viral infections, it is one of the most promising virology aspects. In the present work, a novel broad-spectrum antiviral method, dubbed Double-stranded RNA (dsRNA) Activated Caspase Oligomerizer (DRACO) have been developed, which induces apoptosis in cells with viral dsRNA selectively to kill infected cells quickly with no damage to uninfected ones.

    Materials and Methods

    Following the design, development, expression, and purification of DRACO, influenza virus-infected MDCK and uninfected MDCK cells were treated with 40, 60, and 80 mg/L concentration of DRACO to study its potential antiviral activity. Then, TCID50 (50% Tissue Culture Infectious Dose) of the virus, together with the viability of cells, was measured.

    Results

    The findings of the present study showed that DRACO is nontoxic to uninfected MDCK cells and is effective for H1N1 influenza virus-infected MDCK cells dose-dependently. Also, the infected MDCK cells treated with DRACO have shown a significant reduction in TCID50 compared with the control group.

    Conclusions

    The outcomes suggest that DRACO has potential as a new anti-H1N1 therapeutic drug that its in-vivo antiviral efficacy requires to be examined through a clinical analysis of large quantities of animal models.

    Keywords: Double-Stranded RNA, Caspases, MDCK Cells, Cytotoxicity, Antiviral
  • Mahmoud Gharbavi, Ali Sharafi, Puria Motamed Fath, Saeed Oruji, Hossein Pakzad, Hamidreza Kheiri Manjili * Pages 51-62
    Introduction

     Paclitaxel (PTX), an effective chemotherapeutic drug, is widely used to treat several types of cancers.  However, its use is limited due to poor water solubility resulted in poor bioavailability. One of the main ways to increase the efficacy and endurance of medications depends on the carrier that used for it. In the current study, the microemulsions (ME) were investigated based on Poly [2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (BMA)-co-p-nitrophenyl-oxycrabonyl poly ethylene glycol-methacrylate (ME-ONP)] (PMBN) ability as a carrier for PTX to increase the half-life and its bioavailability. Also, the physicochemical properties of the ME system were evaluated.

    Materials and Methods

     PMBN, tween 80, triacetin, and glycerol were used as the drug carrier, surfactant, oil phase, and co-surfactant, respectively. The hemolytic activity was characterized using the RBC hemolysis assay to evaluate the blood compatibility of the MEs. In addition, the in vitro cytotoxic effect of PMBN-PTX-ME and PTX on MCF-7, 4T1, and HFF-2 cell lines was performed. PI and Annexin-V dyes were used for cell apoptosis.

    Results

    The conductivity of ME evaluated and the results showed 432 to 589 µS/cm. In vitro, the drug release study revealed that PTX has controlled release at different pH levels. Refractive Index (RI) and % transmittance of the MEs remained ranging from 1.43 to 1.53, and 89% to 98%, respectively. The MTT assay determined that PMBN-ME without PTX had no significant toxicity on HFF-2, MCF-7, and 4T-1 cell lines. Based on the apoptosis assay, treated cell lines with PMBN approximately remained alive.

    Conclusions

    The results revealed that ME-based on PMBN would be a promising drug delivery system for PTX drug delivery.

    Keywords: PMBN polymer, Microemulsion, Paclitaxel
  • Asghar Estaji *, Esmaeil Chamani, Zahra Khazaei Pages 63-70
    Introduction

    Lily is often described as one of the most widespread, commercial crops in the floriculture industry. Commercially grown cultivars are mostly propagated by bulb scales which it cost effective and uniformity in tissue culture conditions. Plant tissue culture techniques can effectively provide far-reaching implications in micro-propagation.

    Materials and Methods

    This study was conducted to investigate the effects of various concentrations of cytokines and auxins on callogenesis and Biomass under in vitro conditions. In this experiment, the evaluations were aimed at measuring different characteristics in two lily cultivars, namely, Lilium ledebourii and Lilium regal. 

    Results

    The results showed significant values in all of the measured characteristics. The highest percentage of callogenesis was caused by 2 µm PIC plus 1 µm KIN in L. ledebourii (88.66%) and L. regal (88.66%). Also, the callus weight in both cultivars was obtained by applying the same combination of treatments. In the second experiment, the highest fresh biomass of the cell suspension occurred by applying 2 µm PIC plus 4 µm KIN. The maximum amount of fresh biomass in L. ledebourii (88.93 g/L) and L. regal (41 g/L) occurred on the 24th and 20th day of the culture, respectively.

    Conclusions

     An efficient and fragile callus induction was developed by NAA, which nonetheless made the condition more suitable for cell suspension culture. These lily cultivars need high amounts of PIC as auxin to grow well in cell suspensions. By increasing the PIC level, the biomass accumulates more. These results can moderately optimize large-scale production of both fragile calli and cell suspension biomass.

    Keywords: Callus Formation, cell suspension culture, Lilium
  • Jamal Rashidiani, Khadijeh Eskandari, Reza Ranjbar *, Hamid Kooshki, Davoud Afshar, Farhad Safarpoor Dehkordi Pages 71-75
    Introduction

    For rapid and sensitive detection of Vibrio cholerae an accurate assay is needed. In the present study, a gold-coated magnetic nanoparticle (GMNP) was investigated for the detection of V. cholerae.

    Materials and Methods

    GMNPs were synthesized and functionalized by 11-mercapto-undecanoic acid (MUA) as a linker for immobilization of IgG against V. cholerae OmpW antigen. In the next step, IgG was coupled with carboxylic group of MUA using EDC / NHS and the IgG/GMNPs nanocomposite created and finally, the bacterium was detected in a sandwich model ELISA.

    Results

    The IgG/GMNPs nanocomposite could detect V.cholerae in a concentration range from 2.5×10-2 to 1.5 × 105 N/ml (number of V.cholerae per ml). The correlation coefficient was 0.99 and the detection limit was 16 N/ml.

    Conclusions

    In this study, the procedure of antibody immobilization on magnetic nanoparticles was designed. So that, by using magnetic nanoparticles, the pre-concentration as a time-consuming step was removed and the sensitivity of V. cholerae determination was increased. Also, this method can be extended to detect other microorganisms.

    Keywords: Vibrio cholerae, Gold, Magnetic nanoparticles, Detection
  • Abdollah Ramzani Ghara, Fereshteh Ezzati Ghadi *, Seyed Hamzeh Hossaini, Sonia Poacente, Antonieta Cerulli, Akram Alizadeh, Rouhollah Mirmahmoudi Pages 76-82
    Introduction

    The twig of Capparis decidua has been traditionally used as an antidiabetic agent but its medicinal use has not been scientifically proved yet. Present study evaluated the antidiabetic effect and antioxidant activities of aquatic extract of twigof Capparis decidua on liver and pancreas of diabetic rats induced by Streptozotocin (STZ).

    Materials and Methods

    The effect of Capparis decidua was evaluated by biochemical, histological and FTIR studies. All biochemical and biophysical parameters were estimated after 15 days of daily oral administrations of the aqueous extract.

    Results

    Results showed that oral use of Capparis decidua extract (250 mg/kg) caused significant reduction in the fasting blood glucose levels in diabetic rats when compared to control rats. Moreover, the altered level of lipid peroxidation, antioxidant, lipid profiles, liver enzymes, and also structural changes were reversed in STZ-induced diabetic rats which received Capparis decidua extract.

    Conclusions

    In conclusion, aqueous extract of Capparis decidua has potent antidiabetic and antioxidant activity.

    Keywords: Capparis decidua, Antioxidant, Liver, pancreas, Diabetes