Publications by authors named "Meysam Sarshar"

23 Publications

  • Page 1 of 1

Urinary tract infections: Can we prevent uropathogenic infection with dietary intervention?

Int J Vitam Nutr Res 2021 Apr 21:1-5. Epub 2021 Apr 21.

San Raffaele Roma Open University, Rome, Italy.

Urinary tract infections (UTIs) are among the most common causes of infections in women. Via the fecal-perineal-urethral route, uropathogenic (UPEC) can cause ascending urinary tract infections, including cystitis and pyelonephritis. These infections re-occur within six months or they account for, at least, three episodes within a year of recurrent UTIs (rUTIs). Long term and continuous antibiotic treatment or prophylaxis should be considered as the last options in rUTIs. Conversely, updated European Association of Urology guidelines recommend non-antimicrobial approaches to prevent rUTIs. Accordingly, several studies reported the efficacy of number of natural molecules in inhibiting UPEC adhesion to bladder cells, restraining bacterial growth, as well as stimulating the host innate immune defenses, and protecting the bladder and the kidney mucosa. Therefore, we propose an "anti-UPEC" diet enriched of foods containing natural compounds that were proven effective against UPEC, such as D-mannose, cranberry extracts and medicinal plants. Being a valuable and safe clinical approach to reduce UTI recurrence and limiting the detrimental effects of long and continuous antibiotic prophylaxis, dietary interventions should be evaluated in future clinical trials.
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http://dx.doi.org/10.1024/0300-9831/a000704DOI Listing
April 2021

: An Ancient Commensal with Weapons of a Pathogen.

Pathogens 2021 Mar 24;10(4). Epub 2021 Mar 24.

IRCCS San Raffaele Pisana, 00166 Rome, Italy.

is regarded as a life-threatening pathogen associated with community-acquired and nosocomial infections, mainly pneumonia. The rise in the number of antibiotic-resistant strains reduces effective therapies and increases mortality. Bacterial comparative genomic studies have unraveled the innate and acquired virulence factors of These virulence factors are involved in antibiotic resistance, environmental persistence, host-pathogen interactions, and immune evasion. Studies on host-pathogen interactions revealed that evolved different mechanisms to adhere to in order to invade host respiratory cells as well as evade the host immune system. In this review, we discuss current data on genetic features and virulence factors. An emphasis is given to the players in host-pathogen interaction in the respiratory tract. In addition, we report recent investigations into host defense systems using in vitro and in vivo models, providing new insights into the innate immune response to infections. Increasing our knowledge of pathogenesis may help the development of novel therapeutic strategies based on anti-adhesive, anti-virulence, and anti-cell to cell signaling pathways drugs.
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http://dx.doi.org/10.3390/pathogens10040387DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8063835PMC
March 2021

Acinetobacter baumannii Targets Human Carcinoembryonic Antigen-Related Cell Adhesion Molecules (CEACAMs) for Invasion of Pneumocytes.

mSystems 2020 Dec 22;5(6). Epub 2020 Dec 22.

IRCCS San Raffaele Pisana, Department of Human Sciences and Promotion of the Quality of Life, San Raffaele Roma Open University, Rome, Italy.

Multidrug-resistant is regarded as a life-threatening pathogen mainly associated with nosocomial and community-acquired pneumonia. Here, we show that can bind the human carcinoembryonic antigen-related cell adhesion molecule (CEACAM) receptors CEACAM1, CEACAM5, and CEACAM6. This specific interaction enhances internalization in membrane-bound vacuoles, promptly decorated with Rab5, Rab7, and lipidated microtubule-associated protein light chain 3 (LC3). Dissecting intracellular signaling pathways revealed that infected pneumocytes trigger interleukin-8 (IL-8) secretion via the extracellular signal-regulated kinase (ERK)1/2 and nuclear factor-kappa B (NF-κB) signaling pathways for clearance. However, in CEACAM1-L-expressing cells, IL-8 secretion lasts only 24 h, possibly due to an -dependent effect on the CEACAM1-L intracellular domain. Conversely, the glycosylphosphatidylinositol-anchored CEACAM5 and CEACAM6 activate the c-Jun NH-terminal kinase (JNK)1/2-Rubicon-NOX2 pathway, suggestive of LC3-associated phagocytosis. Overall, our data show for the first time novel mechanisms of adhesion to and invasion of pneumocytes by via CEACAM-dependent signaling pathways that eventually lead to bacterial killing. These findings suggest that CEACAM upregulation could put patients at increased risk of lower respiratory tract infection by This work shows for the first time that binds to carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), CEACAM5, and CEACAM6. This binding significantly enhances internalization within alveolar host cell epithelia. Intracellular trafficking involves typical Rab5 and Rab7 vacuolar proteins as well as light chain 3 (LC3) and slowly progresses to bacterial killing by endosome acidification. CEACAM engagement by leads to distinct and specific downstream signaling pathways. The CEACAM1 pathway finely tunes interleukin-8 (IL-8) secretion, whereas CEACAM5 and CEACAM6 mediate LC3-associated phagocytosis. The present study provides new insights into -host interactions and could represent a promising therapeutic strategy to reduce pulmonary infections caused by this pathogen.
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http://dx.doi.org/10.1128/mSystems.00604-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7762790PMC
December 2020

SARS-CoV-2: Comparative analysis of different RNA extraction methods.

J Virol Methods 2021 01 4;287:114008. Epub 2020 Nov 4.

San Raffaele Roma Open University, 00166 Rome, Italy; IRCCS San Raffaele Pisana, 00166 Rome, Italy.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiologic agent of the COVID-19 pandemic. Although other diagnostic methods have been introduced, detection of viral genes on oro- and nasopharyngeal swabs by reverse-transcription real time-PCR (rRT-PCR) assays is still the gold standard. Efficient viral RNA extraction is a prerequisite for downstream performance of rRT-PCR assays. Currently, several automatic methods that include RNA extraction are available. However, due to the growing demand, a shortage in kit supplies could be experienced in several labs. For these reasons, the use of different commercial or in-house protocols for RNA extraction may increase the possibility to analyze high number of samples. Herein, we compared the efficiency of RNA extraction of three different commercial kits and an in-house extraction protocol using synthetic ssRNA standards of SARS-CoV-2 as well as in oro-nasopharyngeal swabs from six COVID-19-positive patients. It was concluded that tested commercial kits can be used with some modifications for the detection of the SARS-CoV-2 genome by rRT-PCR approaches, although with some differences in RNA yields. Conversely, EXTRAzol reagent was the less efficient due to the phase separation principle at the basis of RNA extraction. Overall, this study offers alternative suitable methods to manually extract RNA that can be taken into account for SARS-CoV-2 detection.
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http://dx.doi.org/10.1016/j.jviromet.2020.114008DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7640895PMC
January 2021

The Global Emergency of Novel Coronavirus (SARS-CoV-2): An Update of the Current Status and Forecasting.

Int J Environ Res Public Health 2020 08 5;17(16). Epub 2020 Aug 5.

Department of Public Health and Infectious Diseases, Sapienza University of Rome, Laboratory affiliated to Institute Pasteur Italia- Cenci Bolognetti Foundation, 00185 Rome, Italy.

Over the past two decades, there have been two major outbreaks where the crossover of animal to humans has resulted in severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV). In December 2019, a global public health concern started with the emergence of a new strain of coronavirus (SARS-CoV-2 or 2019 novel coronavirus, 2019-nCoV) which has rapidly spread all over the world from its origin in Wuhan, China. SARS-CoV-2 belongs to the genus, which includes human SARS-CoV, MERS and two other human coronaviruses (HCoVs), HCoV-OC43 and HCoV-HKU1. The fatality rate of SARS-CoV-2 is lower than the two previous coronavirus epidemics, but it is faster spreading and the large number of infected people with severe viral pneumonia and respiratory illness, showed SARS-CoV-2 to be highly contagious. Based on the current published evidence, herein we summarize the origin, genetics, epidemiology, clinical manifestations, preventions, diagnosis and up to date treatments of SARS-CoV-2 infections in comparison with those caused by SARS-CoV and MERS-CoV. Moreover, the possible impact of weather conditions on the transmission of SARS-CoV-2 is also discussed. Therefore, the aim of the present review is to reconsider the two previous pandemics and provide a reference for future studies as well as therapeutic approaches.
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http://dx.doi.org/10.3390/ijerph17165648DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7459861PMC
August 2020

Fecal microRNAs as Innovative Biomarkers of Intestinal Diseases and Effective Players in Host-Microbiome Interactions.

Cancers (Basel) 2020 Aug 5;12(8). Epub 2020 Aug 5.

Research Laboratories, Bambino Gesù Children's Hospital, IRCCS, 00146 Rome, Italy.

Over the past decade, short non-coding microRNAs (miRNAs), including circulating and fecal miRNAs have emerged as important modulators of various cellular processes by regulating the expression of target genes. Recent studies revealed the role of miRNAs as powerful biomarkers in disease diagnosis and for the development of innovative therapeutic applications in several human conditions, including intestinal diseases. In this review, we explored the literature and summarized the role of identified dysregulated fecal miRNAs in intestinal diseases, with particular focus on colorectal cancer (CRC) and celiac disease (CD). The aim of this review is to highlight one fascinating aspect of fecal miRNA function related to gut microbiota shaping and bacterial metabolism influencing. The role of miRNAs as "messenger" molecules for inter kingdom communications will be analyzed to highlight their role in the complex host-bacteria interactions. Moreover, whether fecal miRNAs could open up new perspectives to develop novel suitable biomarkers for disease detection and innovative therapeutic approaches to restore microbiota balance will be discussed.
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http://dx.doi.org/10.3390/cancers12082174DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7463924PMC
August 2020

FimH and Anti-Adhesive Therapeutics: A Disarming Strategy Against Uropathogens.

Antibiotics (Basel) 2020 Jul 10;9(7). Epub 2020 Jul 10.

Department of Public Health and Infectious Diseases, Sapienza University of Rome, 00185 Rome, Italy.

Chaperone-usher fimbrial adhesins are powerful weapons against the uropathogens that allow the establishment of urinary tract infections (UTIs). As the antibiotic therapeutic strategy has become less effective in the treatment of uropathogen-related UTIs, the anti-adhesive molecules active against fimbrial adhesins, key determinants of urovirulence, are attractive alternatives. The best-characterized bacterial adhesin is FimH, produced by uropathogenic (UPEC). Hence, a number of high-affinity mono- and polyvalent mannose-based FimH antagonists, characterized by different bioavailabilities, have been reported. Given that antagonist affinities are firmly associated with the functional heterogeneities of different FimH variants, several FimH inhibitors have been developed using ligand-drug discovery strategies to generate high-affinity molecules for successful anti-adhesion therapy. As clinical trials have shown d-mannose's efficacy in UTIs prevention, it is supposed that mannosides could be a first-in-class strategy not only for UTIs, but also to combat other Gram-negative bacterial infections. Therefore, the current review discusses valuable and effective FimH anti-adhesive molecules active against UTIs, from design and synthesis to in vitro and in vivo evaluations.
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http://dx.doi.org/10.3390/antibiotics9070397DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7400442PMC
July 2020

Optimization of activin-A: a breakthrough in differentiation of human induced pluripotent stem cell into definitive endoderm.

3 Biotech 2020 May 27;10(5):215. Epub 2020 Apr 27.

6Department of Translational Research and Cellular Therapeutics, Diabetes and Metabolism Research Institute, Beckman Research Institute of City of Hope, Duarte, USA.

The first step in differentiation of pluripotent stem cell toward endoderm-derived cell/organ is differentiation to definitive endoderm (DE) which is the central issue in developmental biology. Based on several evidences, we hypothesized that activin-A optimization as well as replacement of fetal bovine serum (FBS) with knockout serum replacement (KSR) is important for differentiation of induced pluripotent stem cell (iPSC) line into DE. Therefore, a stepwise differentiation protocol was applied on R1-hiPSC1 cell line. At first, activin-A concentration (30, 50, 70 and 100 ng/ml) was optimized. Then, substitution of FBS with KSR was evaluated across four treatment groups. The amount of differentiation of iPSC toward DE was determined by quantitative gene expression analyses of pluripotency ( and ), definitive endoderm ( and ) and endoderm-derived organs (, and ). Based on gene expression analyses, the more decrease in concentrations of activin-A can increase the differentiation of iPSC into DE, therefore, 30 ng/ml activin-A was chosen as the best concentration for the differentiation of R1-hiPSC1 line toward endoderm-derived organ. Moreover, complete replacement of FBS with gradually increased KSR improved the differentiation of iPSC toward DE. For this reason, the addition of 0% KSR at day 1, 0.2% at day 2 and 2% for the next 3 days was the best optimal protocol of the differentiation of iPSC toward DE. Overall, our results demonstrate that optimization of activin-A is important for differentiation of iPSC line. Furthermore, the replacement of FBS with KSR can improve the efficiency of iPSC differentiation toward DE.
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http://dx.doi.org/10.1007/s13205-020-02215-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7186283PMC
May 2020

Protective Effect of HLA-E∗0101∕∗0103 Genotype in Survival of Patients After Allogeneic Hematopoietic Stem Cell Transplant.

Exp Clin Transplant 2020 Apr 7. Epub 2020 Apr 7.

From the Transplant Research Center, Shiraz University of Medical Sciences; and the Islamic Azad University, Arsanjan Branch, Shiraz, Iran.

Objectives: HLA-E is located on the nonclassical major histocompatibility complex class I and acts as the ligand for natural killer cells. Consequently, it has a main role in the regulation of innate immune responses by involving cell identification by natural killer cells. Differences in expression levels among HLA-E alleles have been suggested to affect transplant outcomes. In this study, we evaluated the effects of different HLA-E genotypes on allogeneic hematopoietic stem cell transplant in southern Iran.

Materials And Methods: We investigated 200 patients (donors and recipients) who underwent allogeneic hematopoietic stem-cell transplant and 100 normal participants (control group) in a case-control study. Detection of HLA-E polymorphisms was performed using a sequence-specific primer polymerase chain reaction method.

Results: Statistical analyses indicated that genotypes in the transplant group were not distributed in accordance with Hardy-Weinberg equilibrium (χ²=76.56; P < .001), whereas genotypes in the control group were distributed in accordance with Hardy-Weinberg equilibrium (χ²=0.39; P = .53). No significant differences were observed in cumulative incidence of acute (P = .76; hazard ratio = 0.80; 95% confidence interval, 0.19-3.31) and chronic (P = .75, hazard ratio = 0.048; 95% confidence interval, 0.00) graft-versus-host disease in recipients harboring HLA-E∗0103 allele compared with those homozygous for the HLA-E∗0101 allele. The HLA-E∗0103 allele showed a trend toward lower cumulative incidence of relapse compared with the homozygous HLA-E∗0101 genotype (8% vs 21.5%; P = .37; hazard ratio = 2.50; 95% confidence interval, 0.32-19.20).

Conclusions: Genotypes of the HLA-E molecule did not correlate with acute and chronic graft-versus-host disease in hematopoietic stem cell transplant recipients except for the HLA-E∗0101∗∕∗0103 genotype, which was protective in survival of our study patients.
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http://dx.doi.org/10.6002/ect.2019.0370DOI Listing
April 2020

d-Mannose Treatment neither Affects Uropathogenic Properties nor Induces Stable FimH Modifications.

Molecules 2020 Jan 13;25(2). Epub 2020 Jan 13.

IRCCS San Raffaele Pisana, Department of Human Sciences and Promotion of the Quality of Life, San Raffaele Roma Open University, 00166 Rome, Italy.

Urinary tract infections (UTIs) are mainly caused by uropathogenic (UPEC). Acute and recurrent UTIs are commonly treated with antibiotics, the efficacy of which is limited by the emergence of antibiotic resistant strains. The natural sugar d-mannose is considered as an alternative to antibiotics due to its ability to mask the bacterial adhesin FimH, thereby preventing its binding to urothelial cells. Despite its extensive use, the possibility that d-mannose exerts "antibiotic-like" activity by altering bacterial growth and metabolism or selecting FimH variants has not been investigated yet. To this aim, main bacterial features of the prototype UPEC strain CFT073 treated with d-mannose were analyzed by standard microbiological methods. FimH functionality was analyzed by yeast agglutination and human bladder cell adhesion assays. Our results indicate that high d-mannose concentrations have no effect on bacterial growth and do not interfere with the activity of different antibiotics. d-mannose ranked as the least preferred carbon source to support bacterial metabolism and growth, in comparison with d-glucose, d-fructose, and l-arabinose. Since small glucose amounts are physiologically detectable in urine, we can conclude that the presence of d-mannose is irrelevant for bacterial metabolism. Moreover, d-mannose removal after long-term exposure did not alter FimH's capacity to bind to mannosylated proteins. Overall, our data indicate that d-mannose is a good alternative in the prevention and treatment of UPEC-related UTIs.
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http://dx.doi.org/10.3390/molecules25020316DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7024335PMC
January 2020

A simple, fast and reliable scan-based technique as a novel approach to quantify intracellular bacteria.

BMC Microbiol 2019 11 12;19(1):252. Epub 2019 Nov 12.

Department of Public Health and Infectious Diseases, Sapienza University of Rome, 00185, Rome, Italy.

Background: Quantification of intracellular bacteria is fundamental in many areas of cellular and clinical microbiology to study acute and chronic infections. Therefore, rapid, accurate and low-cost methods represent valuable tools in determining bacterial ability to persist and proliferate within eukaryotic cells.

Results: Herein, we present the first application of the immunofluorescence In-Cell Western (ICW) assay aimed at quantifying intracellular bacteria in in vitro infection models. The performance of this new approach was evaluated in cell culture infection models using three microorganisms with different lifestyles. Two facultative intracellular bacteria, the fast-growing Shigella flexneri and a persistent strain of Escherichia coli, as well as the obligate intracellular bacterium Chlamydia trachomatis were chosen as bacterial models. The ICW assay was performed in parallel with conventional quantification methods, i.e. colony forming units (CFUs) and inclusion forming units (IFUs). The fluorescence signal intensity values from the ICW assay were highly correlated to CFU/IFUs counting and showed coefficients of determination (R), ranging from 0,92 to 0,99.

Conclusions: The ICW assay offers several advantages including sensitivity, reproducibility, high speed, operator-independent data acquisition and overtime stability of fluorescence signals. All these features, together with the simplicity in performance, make this assay particularly suitable for high-throughput screening and diagnostic approaches.
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http://dx.doi.org/10.1186/s12866-019-1625-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6849193PMC
November 2019

Insights into the Periplasmic Proteins of AB5075 and the Impact of Imipenem Exposure: A Proteomic Approach.

Int J Mol Sci 2019 Jul 13;20(14). Epub 2019 Jul 13.

Department of Public Health and Infectious Diseases, Sapienza University of Rome, 00185 Rome, Italy.

Carbapenem-resistant strains cause life-threatening infections due to the lack of therapeutic options. Although the main mechanisms underlying antibiotic-resistance have been extensively studied, the general response to maintain bacterial viability under antibiotic exposure deserves to be fully investigated. Since the periplasmic space contains several proteins with crucial cellular functions, besides carbapenemases, we decided to study the periplasmic proteome of the multidrug-resistant (MDR) AB5075 strain, grown in the absence and presence of imipenem (IMP). Through the proteomic approach, 65 unique periplasmic proteins common in both growth conditions were identified: eight proteins involved in protein fate, response to oxidative stress, energy metabolism, antibiotic-resistance, were differentially expressed. Among them, ABUW_1746 and ABUW_2363 gene products presented the tetratricopeptide repeat motif, mediating protein-protein interactions. The expression switch of these proteins might determine specific protein interactions to better adapt to changing environmental conditions. ABUW_2868, encoding a heat shock protein likely involved in protection against oxidative stress, was upregulated in IMP-exposed bacteria. Accordingly, the addition of periplasmic proteins from cultured with IMP increased bacterial viability in an antioxidant activity assay. Overall, this study provides the first insights about the composition of the periplasmic proteins of a MDR strain, its biological response to IMP and suggests possible new targets to develop alternative antibiotic drugs.
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http://dx.doi.org/10.3390/ijms20143451DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6679007PMC
July 2019

Colonic adenoma-associated Escherichia coli express specific phenotypes.

Microbes Infect 2019 Aug - Sep;21(7):305-312. Epub 2019 Feb 11.

Department of Public Health and Infectious Diseases, Sapienza University of Rome, P.le A. Moro 5, 00185, Rome, Italy; Dani Di Giò Foundation-Onlus, Rome, Italy. Electronic address:

Specific Escherichia coli strains have been associated to colorectal cancer, while no data are available on genotypic and phenotypic features of E. coli colonizing premalignant adenomatous polyps and their pathogenic potential. This study was aimed at characterizing isolates collected from polyps and adjacent tissue in comparison with those from normal mucosa. From colonoscopy biopsies, 1500 E. coli isolates were retrieved and genotyped; 272 were characterized for phylogroup and major phenotypic traits (i.e., biofilm formation, motility, hemolysins, and proteases). Selected isolates were analyzed for extraintestinal pathogenic E. coli (ExPEC)-associated virulence genes and in vivo pathogenicity using Galleria mellonella. The majority of isolates collected from polyps were strong biofilm and poor protease producers, whereas those isolates from normal mucosa were highly motile, proteolytic and weak biofilm formers. Isolates from adjacent tissues shared features with those from both polyps and normal mucosa. Among selected E. coli isolates, ExPEC gene content/profile was variable and uncorrelated with the tissue of collection and larval mortality. Despite the heterogeneous virulence-gene carriage of the E. coli intestinal population, E. coli colonizing colonic adenomatous polyps express specific phenotypic traits that could represent an initial pathoadaptation to local environmental changes characterizing these lesions.
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http://dx.doi.org/10.1016/j.micinf.2019.02.001DOI Listing
February 2020

Genetic diversity, phylogroup distribution and virulence gene profile of pks positive Escherichia coli colonizing human intestinal polyps.

Microb Pathog 2017 Nov 5;112:274-278. Epub 2017 Oct 5.

Department of Public Health and Infectious Diseases, Sapienza University of Rome, Rome, Italy.

Some Escherichia coli strains of phylogroup B2 harbor a (pks) pathogenicity island that encodes a polyketide-peptide genotoxin called colibactin. It causes DNA double-strand breaks and megalocytosis in eukaryotic cells and it may contribute to cancer development. Study of bacterial community that colonizes the adenomatous polyp lesion, defined as precancerous lesions, could be helpful to assess if such pathogenic bacteria possess a role in the polyp progression to cancer. In this cross-sectional study, a total of 1500 E. coli isolates were obtained from biopsies of patients presenting adenomatous colon polyps, the normal tissues adjacent to the polyp lesion and patients presenting normal mucosa. pks island frequency, phylogenetic grouping, fingerprint genotyping, and virulence gene features of pks positive (pks) E. coli isolates were performed. We found pksE. coli strongly colonize two patients presenting polypoid lesions and none were identified in patients presenting normal mucosa. Predominant phylogroups among pksE. coli isolates were B2, followed by D. Clustering based on fragment profiles of composite analysis, typed the pks isolates into 5 major clusters (I-V) and 17 sub-clusters, demonstrating a high level of genetic diversity among them. The most prevalent virulence genes were fimH and fyuA (100%), followed by vat (92%), hra and papA (69%), ibeA (28%), and hlyA (25%). Our results revealed that pksE. coli can colonize the precancerous lesions, with a high distribution in both the polyp lesions and in normal tissues adjacent to the lesion. The high differences in fingerprinting patterns obtained indicate that pksE. coli strains were genetically diverse, possibly allowing them to more easily adapt to environmental variations.
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http://dx.doi.org/10.1016/j.micpath.2017.10.009DOI Listing
November 2017

Microbiota-Derived Extracellular Vesicles as New Systemic Regulators.

Front Microbiol 2017 24;8:1610. Epub 2017 Aug 24.

Mycobacteriology and Pulmonary Research Department, Pasteur Institute of IranTehran, Iran.

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http://dx.doi.org/10.3389/fmicb.2017.01610DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5573799PMC
August 2017

Detection of eight foodborne bacterial pathogens by oligonucleotide array hybridization.

Electron Physician 2017 May 25;9(5):4405-4411. Epub 2017 May 25.

Department of Public Health and Infectious Diseases, Sapienza University of Rome, Laboratory affiliated to Institute Pasteur Italia-Fondazione Cenci Bolognetti, Rome, Italy.

Background: Simultaneous and rapid detection of multiple foodborne bacterial pathogens is important for the prevention of foodborne illnesses.

Objective: The aim of this study was to evaluate the use of and sequences as targets for simultaneous detection of eight foodborne bacterial pathogens.

Methods: Nineteen bacterial oligonucleotide probes were synthesized and applied to nylon membranes. Digoxygenin labeled and from bacteria were amplified by PCR using universal primers, and the amplicons were hybridized to the membrane array. Hybridization signals were visualized by NBT/BCIP color development.

Results: The eight intestinal bacterial pathogens including , , , , , , , and were appropriately detected in a panel of oligonucleotide array hybridization. The experimental results showed that the method could discriminate the bacterial pathogens successfully. The sensitivity of oligonucleotide array was 10 CFU/ml.

Conclusion: This study showed that and genes had sufficient sequence diversity for species identification and were useful for monitoring the populations of foodborne pathogenic bacteria. Furthermore, results obtained in this study revealed that oligonucleotide array hybridization had a powerful capability to detect and identify the bacterial pathogens simultaneously.
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http://dx.doi.org/10.19082/4405DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5498707PMC
May 2017

Simultaneous Molecular Detection of Serovars Typhi, Enteritidis, Infantis, and Typhimurium.

Iran J Public Health 2017 Jan;46(1):103-111

Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.

Background: serovar Typhi, as causative agent of typhoid fever, is one of the most important endemic pathogens. Non-typhoidal serovars, including Typhimurium, Infantis, and Enteritidis are amongst the most prevalent serotypes worldwide and in developing areas such as Iran. The aim of this study was to apply a uniplex PCR for rapid detection of spp., and a multiplex PCR for the simultaneous detection of the four most common serovars in Iran.

Methods: Current research was done in 2010 at Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran. For detection of spp a pair of primers was used to replicate a chromosomal sequence. Four other sets of primers were also designed to amplify the target genes of four species including , and three non-typhoidal spp (). The assay specificity was investigated by testing 15 different serovars and 8 other additional non- species.

Results: The genus-specific PCR yielded the expected DNA band of 404 bp in all spp., strains tested. The uniplex and multiplex PCR assays produced also the expected fragments of 489 bp, 304 bp, 224 bp, and 104 bp for serovars Typhi, Enteritidis, Typhimurium, and Infantis, respectively. Each species-specific primer pair set did not show any cross-reactivity when tested on other serovars or other non- but related- strains.

Conclusion: Both uniplex and multiplex PCR protocols had a good specificity. They can provide an important tool for the rapid and simultaneous detection and differentiation of the four most prevalent serovars in Iran.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5401918PMC
January 2017

Molecular Epidemiology of ESBL Genes and Multi-Drug Resistance in Diarrheagenic Escherichia Coli Strains Isolated from Adults in Iran.

Iran J Pharm Res 2015 ;14(4):1257-62

Department of Public Health and Infectious Diseases, Sapienza University of Rome, Rome, Italy.

Resistance to oxyimino cephalosporins antibiotics in Enterobacteriaceae is primarily done by the extended spectrum β-lactamases (ESBLs). Clear identification of risk factors for ESBLs-producing infections is necessary. Therefore, efficient strategies can be developed to decrease outbreak of these infections. The aim of this study was to determine the antibacterial susceptibility and ESBLs pattern of diarrhogenic Escherichia coli (E. coli) strains isolated from adult patients. In the present study, diarrheogenic E. coli strains were isolated from 54 patients from the University of Medical Sciences hospitals in Shiraz. Antimicrobial susceptibility testing was done by disk diffusion method by CLSI criteria. The presence of bla TEM , bla SHV and bla CTX-M genes was investigated by PCR using designated primers. The prevalence of ESBLs-producer E. coli strains was 12.96%. Antimicrobial resistance testing showed a high resistance to cefexime, trimethoprim-sulfamethoxazole, ampicillin and penicillin. Overall, β-lactamase genes were identified in 52 (96.30%) isolates which were identified as 45 (83.33%) bla TEM, 17 (31.48%) blaSHV and 11 (20.37%) blaCTX-M. ESBLs-producer E. coli is very prevalent in Diarrheogenic strains isolated from adult patients. Also, this study clearly showed that the bla TEM gene for ESBLs-producer E. coli was widespread in Iran.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4673955PMC
December 2015

The study of the oipA and dupA genes in Helicobacter pylori strains and their relationship with different gastroduodenal diseases.

Gastroenterol Hepatol Bed Bench 2015 ;8(Suppl 1):S47-53

Gastroenterology and Liver Diseases Research Centre, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran Iran.

Aim: The purpose of this investigation was to determine the oipA and dupA genes of Helicobacter pylori isolates from west of Iran; Chaharmahalo Bakhtiyari region and find their relationship with the severity of the gastroduodenal diseases.

Background: Helicobacter pylori is an organism responsible for many gastroduodenal diseases. Many studies suggest that genetic diversity in H . pylori virulence factors such as oipA and dupA genes is high among isolates of different geographic regions and may cause more severe diseases.

Patients And Methods: In this cross-sectional study, gastric biopsy specimens were taken from 150 patients suffering from gastroduodenal diseases. The presence of ureC, dupA and oipA genes was tested by polymerase chain reaction (PCR).

Results: Overall, 123 (82%) H. pylori strains were isolated from 150 specimens. dupA gene was detected in 41 (33.33%) H.pylori-positive specimens. There was a reverse correlation between this gene and gastric cancer. The oipA gene was found in 88 (71.54%) samples and statistically there was no association between this gene and gastric disorders. As statistical analyses revealed, the presence of the dupA was more common in isolates with the oipA negative.

Conclusion: Based on our findings, the presence of dupA gene can be considered as a marker for the onset of severe diseases. However, the oipA gene cannot be regarded for prediction of gastroenterology diseases. Meanwhile, extended molecular epidemiology researches in other populations are recommended.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4495424PMC
July 2015

Genetic Analysis of cagA and vacA Genes in Helicobacter Pylori Isolates and Their Relationship with Gastroduodenal Diseases in the West of Iran.

Iran Red Crescent Med J 2013 May 5;15(5):371-5. Epub 2013 May 5.

Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran.

Background: Helicobacter pylori have different virulence factors which are associated with several gastroduodenal diseases; however, this association is variable in different geographical regions. Data of genotypes of Iranian H. pylori isolates are few.

Objectives: The aim of the current study was to investigate the cagA/vacA genotypes of Helicobacter pylori isolates and determine the relationship between these genotypes with respect to different gastric disorders in patients of Chaharmahalo Bakhtiarian.

Materials And Methods: In this cross-sectional study, gastric biopsies were taken from 200 patients with gastrodoudenal diseases. Histopathological features were recognized by specialist. The samples were subjected to PCR for detection and identification of ureC, cagA and vacA genes.

Results: The frequency of the vacA genotypes, sa1/m1, s1a/m1b, s1a/m2, s1b/m1a, s1b/m1b, s1b/m2, s1c/m1a, s1c/m1b, s1c/m2, s2/m1a, s2/m1b and s2/m2 were 27(6.6%), 8(4.3%), 45(28.04%), 7(3.7%), 5(2.5%), 10 (6.1%), 12 (7.4%), 4 (2.5%), 18(11%), 6(3.7%), 0 and 22(13.5%) respectively. The cagA gene was detected in 92% of strains. Based on our findings, it seemed that cagPAI and vacA s1 genotypes were associated with some gastric disorders in patients with H. pylori. In this region, the isolates carrying s1a/m2 were the most prevalent.

Conclusions: We found considerable relationship between s1a/m1a, s1a/m2, s2/m2 and s1c/m1a and some gastric disorders. Further studies about the role of H. pylori virulence factors and gastric disorders were recommended.
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http://dx.doi.org/10.5812/ircmj.3732DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3838643PMC
May 2013

Serogroups, virulence genes and antibiotic resistance in Shiga toxin-producing Escherichia coli isolated from diarrheic and non-diarrheic pediatric patients in Iran.

Gut Pathog 2013 Dec 11;5(1):39. Epub 2013 Dec 11.

Department of Microbiology, Shahrekord Branch, Islamic Azad University, P,O, Box 166, Shahrekord, Iran.

Background: From a clinical perspective, it is important to know which serogroups, virulence genes and antibiotic resistance patterns are present in Shiga toxin-producing Escherichia coli strains in pediatric patients suffering from diarrheic and non-diarrheic infections. This is the first study in Iran that has comprehensively investigated the Shiga toxin-producing Escherichia coli -related infection characteristics in diarrheic and non-diarrheic pediatric patients of 0-60 months of age.

Methods: Two-hundred and twenty four diarrheic and 84 non-diarrheic stool specimens were collected from the Baqiyatallah hospital of Tehran, Iran. The stool samples were cultured immediately and those that were E. coli-positive were analyzed for the presence of antibiotic resistance genes and bacterial virulence factors using PCR. Antimicrobial susceptibility testing was performed using disk diffusion method.

Results: One-hundred and fifty four out of 224 (68.75%) diarrheic stools and 31 out of 84 (36.90%) non-diarrheic stools harbored E. coli. In addition, children in 13-24 month-old age group had the highest incidence of infection with this bacterium (77.63%). A significant difference was found between the frequency of Attaching and Effacing Escherichia coli and Enterohaemorrhagic Escherichia coli (P =0.045). The genes encoding Shiga toxins and intimin were the most commonly detected virulence factors. Among all serogroups studied, O26 (27.04%) and O111 (18.85%) had the highest incidences in the diarrheic and non-diarrheic patients. The incidence of genes encoding resistance against sulfonamide (sul1), gentamicin (aac(3)-IV), trimethoprim (aadA1), cephalothin (blaSHV) and tetracycline (tetA) were 82.78%, 68.03%, 60.65%, 56.55% and 51.63%, respectively. High resistance levels against penicillin (100%), tetracycline (86.88%), gentamicin (62.29%) and streptomycin (54.91%) were observed. Marked seasonality in the serogroup distributions was evident, while STEC infections were more common in summer (P =0.041).

Conclusions: Our findings should raise awareness about antibiotic resistance in diarrheic pediatric patients in Iran. Clinicians should exercise caution when prescribing antibiotics, especially during the warmer months of the year.
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http://dx.doi.org/10.1186/1757-4749-5-39DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3866933PMC
December 2013

Uropathogenic Escherichia coli in Iran: serogroup distributions, virulence factors and antimicrobial resistance properties.

Ann Clin Microbiol Antimicrob 2013 Apr 29;12. Epub 2013 Apr 29.

Department of Microbiology, ShahreKord Branch, Islamic Azad University, P.O. Box: 166, ShahreKord, Iran.

Background: Urinary tract infections (UTIs) are one of the most common bacterial infections with global expansion. These infections are predominantly caused by uropathogenic Escherichia coli (UPEC).

Methods: Totally, 123 strains of Escherichia coli isolated from UTIs patients, using bacterial culture method were subjected to polymerase chain reactions for detection of various O- serogroups, some urovirulence factors, antibiotic resistance genes and resistance to 13 different antibiotics.

Results: According to data, the distribution of O1, O2, O6, O7 and O16 serogroups were 2.43%, besides O22, O75 and O83 serogroups were 1.62%. Furthermore, the distribution of O4, O8, O15, O21 and O25 serogroups were 5.69%, 3.25%, 21.13%, 4.06% and 26.01%, respectively. Overall, the fim virulence gene had the highest (86.17%) while the usp virulence gene had the lowest distributions of virulence genes in UPEC strains isolated from UTIs patients. The vat and sen virulence genes were not detected in any UPEC strains. Totally, aadA1 (52.84%), and qnr (46.34%) were the most prevalent antibiotic resistance genes while the distribution of cat1 (15.44%), cmlA (15.44%) and dfrA1 (21.95%) were the least. Resistance to penicillin (100%) and tetracycline (73.98%) had the highest while resistance to nitrofurantoin (5.69%) and trimethoprim (16.26%) had the lowest frequencies.

Conclusions: This study indicated that the UPEC strains which harbored the high numbers of virulence and antibiotic resistance genes had the high ability to cause diseases that are resistant to most antibiotics. In the current situation, it seems that the administration of penicillin and tetracycline for the treatment of UTIs is vain.
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http://dx.doi.org/10.1186/1476-0711-12-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3651382PMC
April 2013

Study of Helicobacter pylori genotype status in saliva, dental plaques, stool and gastric biopsy samples.

World J Gastroenterol 2012 May;18(17):2105-11

Department of Microbiology, ShahreKord Branch, Islamic Azad University, Shahre Kord 166, Iran.

Aim: To compare genotype of Helicobacter pylori (H. pylori) isolated from saliva, dental plaques, gastric biopsy, and stool of each patient in order to evaluate the mode of transmission of H. pylori infection.

Methods: This cross-sectional descriptive study was performed on 300 antral gastric biopsy, saliva, dental plaque and stool samples which were obtained from patients undergoing upper gastrointestinal tract endoscopy referred to endoscopy centre of Hajar hospital of Shahrekord, Iran from March 2010 to February 2011. Initially, H. pylori strains were identified by rapid urease test (RUT) and polymerase chain reaction (PCR) were applied to determine the presence of H. pylori (ureC) and for genotyping of voculating cytotoxin gene A (vacA) and cytotoxin associated gene A (cagA) genes in each specimen. Finally the data were analyzed by using statistical formulas such as Chi-square and Fisher's exact tests to find any significant relationship between these genes and patient's diseases. P < 0.05 was considered statistically significant.

Results: Of 300 gastric biopsy samples, 77.66% were confirmed to be H. pylori positive by PCR assay while this bacterium were detected in 10.72% of saliva, 71.67% of stool samples. We were not able to find it in dental plaque specimens. The prevalence of H. pylori was 90.47% among patients with peptic ulcer disease (PUD), 80% among patients with gastric cancer, and 74.13% among patients with none ulcer dyspepsia (NUD) by PCR assay. The evaluation of vacA and cagA genes showed 6 differences between gastric biopsy and saliva specimens and 11 differences between gastric and stool specimens. 94.42% of H. pylori positive specimens were cagA positive and all samples had amplified band both for vacA s and m regions. There was significant relationship between vacA s1a/m1a and PUD diseases (P = 0.04), s2/m2 genotype and NUD diseases (P = 0.05). No statically significant relationship was found between cagA status with clinical outcomes and vacA genotypes (P = 0.65). The evaluation of vacA and cagA genes showed 6 differences between gastric biopsy and saliva specimens and 11 differences between gastric and stool specimens.

Conclusion: Regard to high similarity in genotype of H. pylori isolates from saliva, stomach and stool, this study support the idea which fecal- oral is the main route of H. pylori transmission and oral cavity may serve as a reservoir for H. pylori, however, remarkable genotype diversity among stomach, saliva and stool samples showed that more than one H. pylori genotype may exist in a same patient.
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http://dx.doi.org/10.3748/wjg.v18.i17.2105DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3342610PMC
May 2012