Publications by authors named "Maryam Sadrnia"

11 Publications

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and evaluation of antibacterial and anti-biofilm properties of five ethnomedicinal plants against oral bacteria by TEM.

Avicenna J Phytomed 2021 Mar-Apr;11(2):180-189

Department of Biology, Arak Branch, Islamic Azad University, Arak, Iran.

Objective: The aim of the present study was to investigate antibacterial and antibiofilm activity of a few medicinal plants against oral bacteria.

Materials And Methods: , , and were extracted. Isolates from oral cavity were identified by microbiological and molecular methods. Minimum inhibitory concentration and minimum bactericidal concentration were determined by Broth microdilution method. The anti-biofilm activity of essential oils and extracts investigated and as a mixture by Broth dilution method. Toxicity of the herbal mixture was assayed by in Wistar rats treated with intradermal injection. Wound healing properties of the herbal mixture against infected wounds on the back of the rats were investigated. Anti-biofilm activity was investigated on tooth surfaces. Bacterial structure changes and fine- structure study were performed by light microscopy and Transmission electron microscopy.

Results: The lowest MIC and MBC for the plant mixtures was 0.0002 mg/ml belonged to and the highest values (0.025 mg/ml) belonged to . The essential oils of , and but not and extracts, were able to remove the biofilms created by the studied bacteria. The herbal mixture was able to completely heal the wound skin of rats in 21 days (p<0.05 compared to control). The mixture was able to decompose the teeth biofilm in 45 seconds. The results of light and electron microscopy showed that the bacterial structure exposed to the herbal mixture was deformed.

Conclusion: It was concluded that the essential oils of and had significant effects on inhibition of oral bacteria biofilm formation.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8051319PMC
April 2021

Comparison of antibacterial effects of a carrier produced in microemulsion system from aqueous extract of Aloe vera with selected antibiotics on Enterobacteriacea.

Iran J Microbiol 2018 Oct;10(5):334-341

Infectious Diseases Research Center (IDRC), Arak University of Medical Sciences, Arak, Iran.

Background And Objectives: Antibiotics resistance has recently increased. The aim of this study was the evaluation of antibacterial efficacy of Aloe vera carrier produced in microemulsion system in comparison with ordinary antibiotics against some Enterobacteriacea.

Materials And Methods: The aquatic extract of Aleo vera was produced by the Soxhlet method and a nonocarrier in the microemulsion system was prepared by two emulsifiers. The clinical isolates of and were obtained from patients and were identified by microbiological methods. Diffusion disk was used for evaluation of antibacterial properties in comparison with selected ordinary antibiotics. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) for tested materials were determined using MTT in the Micro Broth dilution method.

Results: The results proved that effect of carrier on studied isolates is dependent on concentration level. The inhibitory effect of carrier in concentration of 15 μg/ml by 18 mm zone of inhibition for was comparable to Ceftazidime and Cefalothin. The lowest MIC and MBC determined by the Microbroth dilution method with MTT belonged to as 0.1 and 3 μg/ml and higher concentrations belonged to at 7 and 15 μg/ml. The greatest effect of carrier of Aleo vera aquatic extract was observed for and the lowest effect belonged to and

Conclusion: It was concluded that the carrier of Aloe vera produced in microemulsion system was most effective and had equal effects in comparison with ordinary antibiotics against Enterobacteriacea.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6339994PMC
October 2018

Novel Mutations in Gene of Pyrazinamide Resistant Clinical Isolates of .

Sci Pharm 2018 Apr 16;86(2). Epub 2018 Apr 16.

Infectious Diseases Research Center (IDRC), Arak University of Medical Sciences, Arak, 3819693345, I.R. of Iran.

In clinical isolates of (MTB), resistance to pyrazinamide occurs by mutations in any positions of the gene (NC_000962.3) especially in nucleotides 359 and 374. In this study we examined the gene sequence in clinical isolates of MTB. Genomic DNA of 33 clinical isolates of MTB was extracted by the Chelex100 method. The polymerase chain reactions (PCR) were performed using specific primers for amplification of 744 bp amplicon comprising the coding sequences (CDS) of the gene. PCR products were sequenced by an automated sequencing Bioscience system. Additionally, semi Nested-allele specific (sNASP) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods were carried out for verification of probable mutations in nucleotides 359 and 374. Sequencing results showed that from 33 MTB clinical isolates, nine pyrazinamide-resistant isolates have mutations. Furthermore, no mutation was detected in 24 susceptible strains in the entire 561 bp of the gene. Moreover, new mutations of G→A at position 3 of the gene were identified in some of the resistant isolates. Results showed that the sNASP method could detect mutations in nucleotide 359 and 374 of the gene, but the PCR-RFLP method by the SacII enzyme could not detect these mutations. In conclusion, the identification of new mutations in the gene confirmed the probable occurrence of mutations in any nucleotides of the gene sequence in resistant isolates of MTB.
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http://dx.doi.org/10.3390/scipharm86020015DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6027673PMC
April 2018

Insights into Pyrazinamidase and DNA Gyrase Protein Structures in Resistant and Susceptible Clinical Isolates of .

Tanaffos 2016 ;15(3):147-153

Infectious Diseases Research Center (IDRC), Arak University of Medical Sciences, Arak, Iran.

Background: Mutations in and genes cause pyrazinamide (PZA) and fluroquinolone resistance in (). In the present study, structures of pyrazinamidase (PZase) and DNA gyrase proteins were studied in resistant and susceptible clinical isolates of

Materials And Methods: Sixty clinical isolates of were used in this study. Polymerase chain reaction (PCR) amplification of and genes was accomplished on purified DNA. Sequence of the fragments was determined by an Applied BiosystemsTM apparatus. Bioinformatic analysis was performed by online software and three-dimensional (3D) structures of proteins was predicted using Molegro Virtual Docker (MVD) Modeler software.

Results: Amplified 744 and 194 bp fragments of and genes, respectively were yielded suitable sequence results. Predicted 3D structures of proteins showed some differences between wild-type and mutant structures. Mutation in amino acid No.31 (T92C) caused an increase in distance from metal ion position to enzyme active site, but it was considered as a polymorphism. Docking results by MVD revealed a relationship in quinolone resistance-determining regions (QRDR) amino acids in interaction with antibiotic. T92C mutation in PZase from non-polar aliphatic amino acid Ile (ATC) to polar aliphatic amino acid threonine (ACC) was a polymorphism.

Conclusion: Structural changes in two important proteins related to drug resistance were proven in clinical isolates of .
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5304958PMC
January 2016

Transmission Electron Microscopy of XDR Mycobacterium tuberculosis Isolates Grown on High Dose of Ofloxacin.

Sci Pharm 2017 Feb 2;85(1). Epub 2017 Feb 2.

Infectious Diseases Research Center (IDRC), Arak University of Medical Sciences, Arak 3848176941, Iran.

The aim of the study was to investigate behavior of resistant Mycobacterium tuberculosis (MTB) isolates under a high dose of ofloxacin and its morphological changes. 19 extensively drug resistant (XDR) clinical isolates of MTB were grown on Löwenstein-Jensen medium containing progressively increasing concentrations of ofloxacin (2, 4, 8, 16, 32 mg/L). Ultra-structure analyses of resistant isolates grown on ofloxacin were conducted by transmission electron microscopy (TEM). Fixation was carried out by 4% glutaraldehyde in 0.1 M sodium cacodylate buffer on 300 mesh carbon formvar copper grid. The samples were negatively stained with uranium acetate suspension. All19XDRMTBisolatesweregrownandformedcoloniessuccessfullyon2,4,8mg/L,sevenisolates on16mg/L,andfourisolateson32mg/Lofloxacin. Morphologicalchangesandunusualformswere detected in 8, 16 and 32 mg/L ofloxacin at 43%, 76.5% and 81% of cells, respectively. Swollen form (protoplast like), ghost-like cell, degraded forms, and in a few cases, detached cytoplasm from cell wall were clearly detected in high drug concentrations in comparison to control. Changes in morphology were increased with increasing ofloxacin concentrations (p < 0.05). Some XDR isolates could be successfully grown on high doses of ofloxacin (32 mg/L), but with changes in morphology. It was concluded that several magnitudes of the drug doses could not prevent growth of drug resistant forms.
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http://dx.doi.org/10.3390/scipharm85010003DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5387365PMC
February 2017

Minor Contribution of inhA-15 Mutations to the Rapid Detection of Isoniazid Resistance in Mycobacterium Tuberculosis Isolates.

Iran J Med Sci 2016 Mar;41(2):161-3

Medical Laboratory Sciences and Research Center for TB and Pulmonary Diseases, Tabriz University of Medical Sciences, Tabriz, Iran.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4764969PMC
March 2016

Rapid detection of coliforms in drinking water of Arak city using multiplex PCR method in comparison with the standard method of culture (Most Probably Number).

Asian Pac J Trop Biomed 2014 May;4(5):404-9

Tuberculosis and Infectious Research Center and Department of Microbiology, Arak University of Medical Sciences, Iran.

Objective: To analyse molecular detection of coliforms and shorten the time of PCR.

Methods: Rapid detection of coliforms by amplification of lacZ and uidA genes in a multiplex PCR reaction was designed and performed in comparison with most probably number (MPN) method for 16 artificial and 101 field samples. The molecular method was also conducted on isolated coliforms from positive MPN samples; standard sample for verification of microbial method certificated reference material; isolated strains from certificated reference material and standard bacteria. The PCR and electrophoresis parameters were changed for reducing the operation time.

Results: Results of PCR for lacZ and uidA genes were similar in all of standard, operational and artificial samples and showed the 876 bp and 147 bp bands of lacZ and uidA genes by multiplex PCR. PCR results were confirmed by MPN culture method by sensitivity 86% (95% CI: 0.71-0.93). Also the total execution time, with a successful change of factors, was reduced to less than two and a half hour.

Conclusions: Multiplex PCR method with shortened operation time was used for the simultaneous detection of total coliforms and Escherichia coli in distribution system of Arak city. It's recommended to be used at least as an initial screening test, and then the positive samples could be randomly tested by MPN.
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http://dx.doi.org/10.12980/APJTB.4.2014C896DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3985057PMC
May 2014

Study of carD gene sequence in clinical isolates of Mycobacterium tuberculosis.

Acta Microbiol Immunol Hung 2014 Mar;61(1):1-10

Arak University of Medical Sciences Research Center of Molecular Medicine Arak Iran.

Mycobacterium tuberculosis growth rate is closely coupled to rRNA transcription which is regulated through carD gene. The aim of this study was to determine the sequence of carD gene in drug susceptible and resistant clinical isolates of M. tuberculosis and designing of a PCR assay based on carD sequence for rapid detection of this bacterium.Specific primers for amplification of carD gene were carefully designed, so that whole sequence of gene could be amplified; therefore primers were positioned at the upstream (promoter of this gene and ispD gene) and downstream (in ispD gene). DNA from 41 clinical isolates of M. tuberculosis with different pattern of drug resistance was used in the study. PCR conditions and annealing temperature were designed by means of online programs. PCR products were sequenced by ABI system.PCR product of carD gene was a 524 bp fragment. This method could detect all resistant and susceptible strains of M. tuberculosis. The size of amplified fragment was similar in all investigated samples. Sequence analysis showed that there was similar sequence in all of our isolates therefore probably this gene is considered to be conservative. Translation of nucleotide mode to amino acids was showed that TRCF domain in N-terminal of protein CarD was found to be fully conservative.This is the first study on the sequence of carD gene in clinical isolates of M. tuberculosis. This conservative gene is recommended for use as a target for designing of suitable inhibitors as anti-tuberculosis drug because its importance for life of MTB. In the other hand, a PCR detection method based on detection of carD gene was recommended for rapid detection in routine test.
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http://dx.doi.org/10.1556/AMicr.61.2014.1.1DOI Listing
March 2014

Rapid and simple approach for identification of Mycobacterium tuberculosis and M. bovis by detection of regulatory gene whiB7.

Acta Microbiol Immunol Hung 2011 Mar;58(1):65-74

Arak University of Medical Sciences, Tuberculosis and Pediatric Infectious Diseases Research Center, Arak Iran.

Identification of Mycobacterium tuberculosis and M. bovis is necessary for the application of adequate drug therapy. PCR amplification is a good tool for this purpose, but choosing proper target is of a great concern. We describe a PCR assay for fast detection of M. tuberculosis and M. bovis.As a BLAST and BLASTP search we selected regulatory gene whiB7 that encodes multi-drug resistance in this bacterium. Thirty clinical isolates of M. tuberculosis were sequenced and all the mutations in gene whiB7 were detected. The best set of several pairs of primers was selected and used in comparison by rpoB gene for differentiation of M. bovis, M. avium, M. kansasii, M. phlei, M. fortuitum, M. terrae, seven non-pathogenic Mycobacterium isolates and 30 clinical isolates of M. tuberculosis.It was proved that only clinical isolates of M. tuberculosis and M. bovis have positive bands of 667 bp whiB7. Other non-tuberculous and non-pathogenic isolates did not show any positive sign. Furthermore, 667-bp PCR products of whiB7 gene were observed for ten positive sputum samples (preliminarily approved to be positive for M. tuberculosis by commercially real-time based method), but no bands were detected in 5 negative sputum samples. RpoB gene could not differentiate non-tuberculous strains and non-pathogenic isolates from pathogenic clinical isolates. We concluded that PCR amplification of the gene coding for the WhiB7 protein could be successfully used as a good tool for rapid identification of M. tuberculosis and M. bovis. We propose application of this method as a rapid and simple approach in mycobacteriological laboratories.
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http://dx.doi.org/10.1556/AMicr.58.2011.1.7DOI Listing
March 2011

Prevalence of mutations at codon 463 of katG gene in MDR and XDR clinical isolates of Mycobacterium tuberculosis in Belarus and application of the method in rapid diagnosis.

Acta Microbiol Immunol Hung 2011 Mar;58(1):51-63

Arak University of Medical Sciences, Tuberculosis and Pediatric Infectious Diseases Research Center, Arak Iran.

Isoniazid (INH) is a central component of drug regimens used worldwide to treat tuberculosis. In respect to high GC content of Mycobacterium tuberculosis, nonsynonymous mutations are dominant in this group. In this study a collection of 145 M. tuberculosis isolates was used to evaluate the conferring mutations in nucleotide 1388 of katG gene (KatG463) in resistance to isoniazid. A PCR-RFLP method was applied in comparison with DNA sequencing and anti-mycobacterial susceptibility testing. From all studied patients, 98 (67.6%) were men, 47 (32.4%) were women, 3% were <15 and 9% were >65 years old; male to female ratio was 1:2.4. PCR result of katG for a 620-bp amplicon was successful for all purified M. tuberculosis isolates and there was no positive M. tuberculosis culture with PCR negative results (100% specificity). Subsequent PCR RFLP of the katG identified mutation at KatG463 in 33.3%, 57.8% and 59.2% of our clinically susceptible, multidrug resistant TB (MDR) and extensively drug resistant (XDR) isolates, respectively. Strains of H37Rv and Academic had no any mutations in this codon. M. bovis was used as a positive control for mutation in KatG463. Automated DNA sequencing of the katG amplicon from randomly selected INH-susceptible and resistant isolates verified 100% sequence accuracy of the point mutations detected by PCR-RFLP. We concluded that codon 463 was a polymorphic site that is associated to INH resistance (a missense or "quiet" mutation). RFLP results of katG amplicons were identical to those of sequence method. Our PCR-RFLP method has a potential application for rapid diagnosis of M. tuberculosis with a high specificity.
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http://dx.doi.org/10.1556/AMicr.58.2011.1.6DOI Listing
March 2011