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Aim and back ground: In recent years the threat of acquisition of antibiotics resistance caused by over use of antibiotics is growing. The purpose of this study was to survey the pattern of antibiotic resistance and evaluate of Multi-drug resistance Escherichia coli and analyze the plasmid profile of these clinical strains.

Materials and Methods: This study was performed on 83 clinical Escherichia coli, obtained from four hospital (imam Khomeini, Alborz, Kasra, qaem) and two private laboratories (Razi and Kavosh), of Karaj city. Isolates were identified as Escherichia coli based on standard biochemical tests. The isolates were screened for Antimicrobial susceptibility test by 20 antibiotics using Kirby-Bauer disk diffusion method. Plasmid profiles of these isolates have been analyzed by alkaline lysis method.

Result: Antibiotic resistance pattern were observed as follows: Nitrofurantoin 8.4%, Tetracycline 63.8%, Amoxicillin 83%, Ofloxacin 30%, Levofloxacin 30%, Co-trimoxazole 59%, Amikacin 18%, Imipenem 25.3%, Choloramphenicol 16.9%, Cephalexin 61.4%, Gentamycin 28.2%, Norfloxacin 33.7%, Nalidixic acid 56.6%, Cephalothin 48.2%, Ciprofloxacin 32.5%, Ceftazidime 27.7%, Ceftriaxone 37.3%, Ceftizoxime 16.9%, Cefotaxime 38.5%, Amoxycillin/Clavulanic acid 68.7%. Of 83 isolates 92.77% (77) isolates were resistance to more than two unrelated drugs. Of 83 isolates, 65 isolates, (78.3%) showed the presence of plasmid.

Conclusion: The results of this study indicate a growing resistance pattern to the first line antibiotics, prescribed for urinary tract infection. High presence of plasmids in these strains, play an important role in multidrug resistance in these bacteria.

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Aim and Background:
 Pseudomonas aeruginosa is one of the most important pathogen cause infection in hospital in world. Different virulence gene exsist in this bacteria, which interfere in pathogenesis and infection.
 The aim of this study was evaluation of antibiotic susceptibility pattern and identify virulence genes plcH, lasB, plcN of P. aeruginosa isolated from clinical samples.
 
Material and methods:
 In this study 100 Pseudomonas aeruginosa isolated of human cases of burn in 1395.
Antibiotic susceptibility test was performed by disk diffusion method. Genomic DNA were extracted from isolated strains and Multiplex- PCR performed to determine the preavalence  of each of the tested genes.
Result: The scope age of  patients were 20-90 years, and 46 patient were male and 52 were female.The prevalence of plcN and plcH  observed  98 percent and lasB 100 percent.  The highest resistance to antibiotics were observed in  ampicillin and cefoxitin with the frequency of  93% and 96% respectively.
Conclusion:The  high prevalence of plcH, lasB, plcN gene in Pseudomonas aeruginosa isolated of skin case of burn revealed that this genes are an important virulence factors in burn infections.

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Background and Objective:
The most frequently used sources of more stable enzymes are thermophilic bacteria. In recent years microbial L-asparaginase has been widely used as therapeutic agent in the treatment of human cancers. Microbial L-glutaminase is currently used in the treatment of acute lymphoblastic leukemia (ALL) and human immunodeficiency virus (HIV) because of its potential applications in medicine . The aim of present study was to screen new potent thermophilic bacterial isolates producing L-asparaginase and L-glutaminase.
 Methods:
Larijan hotspring was chosen as the location for isolation of the thermophilic bacteria.  Culture and morphological and biochemical tests, were performed. All the bacterial isolates were screened in ortherto L-asparaginase and L-glutaminase  enzyme  activity  using  Rapid Plate assay method. M9  modified media incorporated L-asparaginase or L-glutaminase and phenol red as indicator were used. Thermophilic strains were later identified by 16SrRNA gene ribotyping followed by PCR .
 Results:
 A total of six isolates recovered from Larijan hotspring were found to belong to Bacillus licheniformis. The L-asparaginase activity was observed in LT-1, LT-2, LT-3, LT-4, LT-5 strains.The L-glutaminase produced by Bacillus licheniformis LT-1, LT-3, LT-5 and LT-6 strains. Three thermophilic LT-1, LT-3, LT-5 strains with L-glutaminase and L-asparaginase activity have been successfully isolated.
Conclusion:
Larijan hot spring is one of the potential sources of useful microbial products that can be used as a source of useful different biological products like enzymes.

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The covid-19 virus is a severe and contagious viral infection that appeared in Wuhan, China in late December 2019 and spread rapidly around the world. Despite numerous studies on the new virus, its origins remain unclear. Determining the exact source of this virus can be helpful in finding solutions to control this virus and similar viruses in the future.

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Aim and Background: Human papillomaviruses (HPVs) especially types 16 and 18 are known as the major causes of cervical cancer. Among HPV proteins, L1 and L2 capsid proteins, and also E7 oncoprotein are proposed as target antigens for vaccine design. In the recent years, the recombinant multiepitope polypeptides have attracted a special interest. The goal of this study is the design of L1-L2-E7 fusion construct and evaluation of its expression in bacterial expression system. 
Materials and Methods: In this study, the immunogenic and conserved epitopes of HPV16/18 L1, L2 and E7 proteins were selected using different bioinformatics analyses. After synthesis of the designed L1-L2-E7 fusion sequence in pUC57 cloning vector, its subcloning was performed in pET24a (+) prokaryotic expression vector using EcoRI/ HindIII restriction enzymes. The expression of the recombinant multiepitope polypeptide was done in E.coli Rosetta strain using IPTG inducer, and confirmed by SDS-PAGE and western blotting using anti-His-tag antibody. The expression was optimized under different conditions such as optical density (OD in wavelength of 600 nm), temperature and time after induction, and IPTG concentration.   
Results: The recombinant pET-L1-L2-E7 vector was confirmed by the presence of a clear band (~525 bp) related to the L1-L2-E7 gene on agarose gel after enzyme digestion. The expression of L1-L2-E7 polypeptide in bacteria showed the presence of a clear band (~20 kDa) in SDS-PAGE and western blotting. The best conditions for expression of the recombinant polypeptide were at temperature of 37^◦C, optical density of 0.7-0.8, IPTG concentration of 1mM, and time of 16 h after induction.  
Conclusion: The successful expression of the L1-L2-E7 multiepitope polypeptide was performed in bacterial system. In the next step, the recombinant polypeptide will be purified to use as a vaccine candidate. 

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Aim and Background: The use of plants as medicine has always been considered. In the present study, the effect of aqueous extract collected in spring and autumn of Artemisia aucheri on the survival of HSV- 1 virus-infected cells was investigated.
Materials and Methods: In this study, the effect of aqueous extract of Artemisia aucheri in spring and autumn on the survival of Vero cell line after HSV- 1 infection at different times and concentrations was compared using TCID50 and MTT methods. The effect of aqueous extract of Artemisia aucheri at concentrations of 25, 50, 75, and 100 mg / ml on Vero cell line infected with HSV- 1 virus was performed for 24 hours in DMEM medium containing 2% (FBS) and then a dose of 50% Tissue Culture Infectious Dose(TCID50) was determined. Cell viability was also assessed by MTT assay at 2-, 0, 4, 8, and 24 hours.
Results: The dose of 50% TCID50 with different dilutions of spring and autumn aqueous extracts of Artemisia aucheri was determined on Vero cells infected with (MOI = 0.1) HSV- 1 virus by inverted microscope and compared by MTT method. Concentrations of 50, 75, µg/ml and 100 µg/ml of spring extract and concentrations of 50 and 75 µ g / ml of autumn extract significantly reduced viral infection. Among them, spring extract with a concentration of 75 µ g / ml showed the highest viral inhibition and cell survival. Also, a significant difference was observed between spring and autumn aqueous extracts of Artemisia aucheri on cell survival (r = 0.97, P =0.0002).
Conclusion: The results indicated that the spring aqueous extract of Artemisia aucheri has a significant anti-HSV- 1 antiviral effect compared to the autumn aqueous extract of Artemisia aucheri (A = 0.0001) (P = 0.0001).

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Aim and Background: Rhamnolipids are one of the most important bio surfactants of glycolipids produced by Pseudomonas aeruginosa. In this study, the expression of bio surfactant producing proteins of Pseudomonas aeruginosa in the presence of sodium dodecyl sulfate coated iron nanostructure (Fe/SDS) were evaluated.
Materials and Methods: The expression of Surfactant Protein A enzyme was investigated by SDS-PAGE and Western blot. The enzyme rhamnosyl transferase was analyzed on SDS-PAGE gel and confirmed by Western blotting. The presence of nanoparticles on the surface of bacteria and the morphology of nanoparticles were investigated by TEM and SEM.
Results: According to SEM, nanoparticles with a size of 20 nm and a spherical shape were seen. Binding of Fe / SDS nanoparticles to bio surfactant was confirmed by TEM. The enzyme rhamnosyl transferase with a molecular weight of 40 kDa was expressed in the test sample. The results of protein expression showed that the confirmed of rhamnosyl transferase enzyme increased significantly under the influence of Fe/SDS nanoparticles and showed the confirmed compared to other nanoparticles and the control sample. While SDS and Fe nanoparticles alone are not effective in confirmed this protein and are almost identical to the control sample. Western blotting results of SDS-PAGE confirmed the expression of rhamnolipid transferase under the influence of Fe/SDS nanoparticles.
Conclusion: The results of the present study indicate the importance of Fe/SDS nanoparticles in increasing the expression of rhamnolipid transferase enzyme in the production of bio surfactants from Pseudomonas aeruginosa.
 

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Aim and background: Cancer is mainly caused by defect in natural regulation of programmed cell death (PCD). On the other hand, it is proved that, the EGFR is overexpressed on the surface of many malignant cell. Therefore, immunotoxin against EGFR, which consisting an antibody or fragment of an antibody linked to the toxic moiety is one of the most reliable strategies for cancer treatment. Here we evaluate the effect of a novel immunotoxin molecule harboring ricin, as a toxic moiety on the mRNA level of two main apoptotic molecules on cancerous cell lines.

Materials and method: The Effects of immunotoxin molecule and it’s components including ricin toxin and antibody fragment on the mRNA level of pro- apoptotic BAX and anti-apoptotic BCL-2 and EGF receptor genes on the breast MDA-MB-468 and colorectal HCT-116 cancer cell lines were assayed by qRT-PCR. We used glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as a reference gene.

Results: Treatment with immunotoxin significantly increased the expression of BAX gene and decreased the expression of EGFR and BCL-2 genes (P<0.05) and induced apoptosis. Also, ricin toxin caused a significant decrease in BCL-2 and no significant increase in BAX in cancer cells and decreased EGFR only in breast cancer cells. The scFv part had no significant changes on genes expression.

Conclusion: The present findings showed that the recombinant Panitumumab-Ricin protein, may be an appropriate candidate for the treatment of EGFR overexpressed cancers by induction in apoptosis pathway.

 

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Aim and Background: Rheumatoid arthritis(RA) is a complex and chronic disease of joint inflammation. Environmental factors that are important in genetically predisposed people are involved in the occurrence of the disease and its progress. This disease has a lot to related to genetics, including human class 2 antigens. In various studies, this relation is about 60%. This relationship is more positive in people with Anti-citrullinated protein antibodies (ACPA) than negative ones. The aim of this study is to investigate the relationship between cytokines and soluble proteins in the body of a patient with Rheumatoid arthritis before symptoms appear.
Materials and Methods: In this research, analysis with Enrichr, WebGestalt, and GeneMANIA was used to find genes related to the disease. In order to check the amount of cytokines predicted, ELISA and Real time RT-PCR methods were applied. Results: By examining the System Biology methods, the candidate genes included IL-6R, IL-6 and IL-23A. The samples were taken from 46 RA patients referred to the rheumatology department of the Babol hospital, and a significant decrease (Pv=0.1255) of IL-6 was observed in patients undergoing treatment and a significant increase (Pv=0.4414) in the amount of this cytokine receptor in the serum compared to healthy individuals. While the expression of IL6 and IL6R genes increased in the patient group compared to the healthy group.
Conclusion: Cytokine IL-6 as an inflammatory cytokine in treated patients decreased in serum and showed a significant increase in RNA level.

 

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Aim and Background: Graphene quantum dots (GQDs) conjugated with folic acid have a very high potential to identify cancer cells with high expression of folate receptor and cause tumor death by binding to an anti-cancer agent. The aim of this research is to design and evaluate a new nano-drug delivery system for breast cancer treatment.

Materials and Methods: GQDs nanoparticles were synthesized by pyrolysis of L-glutamic acid and its surface was coated with PEG and amine agent to create a longer half-life and more fluorescence intensity, and it was conjugated with folic acid for better binding to MCF-7 cancer cell line with folic acid receptor by thermal method. The structure of the nanocarrier was investigated using FTIR spectroscopy, fluorimetry, and TEM imaging methods. Penetration of nanoparticles into cells and their fluorescence emission were investigated by confocal microscopy.

Results: The synthesized GQDs had a uniform, relatively spherical structure and a size below 10 nm. With the addition of amine and PEG agents to GQDs nanoparticles, significant fluorescence intensities of up to 1000 a.u. were observed. According to the FTIR results, folic acid was successfully conjugated on the GQDs nanocarrier after using thermal reaction. The entry and fluorescence emission of nanoparticles into MCF-7 cells was confirmed using confocal microscopy.

Conclusion: According to the results, replacing the thermal method in the synthesis of targeted GQDs nanoparticles, instead of using chemical reagents, can provide a cost-effective and easier method. Pegylated and folated GQDs were suitable nanocarriers for targeted drug delivery to the MCF-7 breast cancer cell line.