Publications by authors named "Zahra Meshkat"

97 Publications

Development of detection methods for the diagnosis and analysis of highly toxic metal phosphides: A comprehensive and critical review.

Biotechnol Appl Biochem 2021 May 14. Epub 2021 May 14.

Department of Medical Biotechnology and Nanotechnology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Metal phosphides, especially aluminum phosphide, and phosphine (PH ) are widely used as insecticides and rodenticides for protection of grains during process of storage and transportation. The main reason of poisoning with this compound is related to the conscious ingestion of salts or accidental inhalation of PH . So the early and accurate diagnosis of poisoning can significantly help to the effective clinical treatment or recognition of death cause. PH is somewhat unstable due to reaction with oxygen or hemoglobin leading to formation of oxy-acids phosphorous. Here, we critically reviewed the literature introducing the quantitative and qualitative methods for the detection of metal phosphides, PH , and its products. This study obviously demonstrates that during past years, different diagnosis methods have been remarkably progressed. Head-space gas chromatography and confirmatory colorimetric methods have been as the most popular techniques. Also, the gas sensors are a promising method that must be more progressed.
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http://dx.doi.org/10.1002/bab.2190DOI Listing
May 2021

pulmonary infection in a vitamin D-deficient patient: A case report.

Clin Case Rep 2021 Mar 1;9(3):1146-1149. Epub 2021 Feb 1.

Antimicrobial Resistance Research Center Mashhad University of Medical Sciences Mashhad Iran.

Closer attention should be paid to vitamin D status in patients with mycobacterial diseases.
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http://dx.doi.org/10.1002/ccr3.3692DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7981631PMC
March 2021

Immunogenicity of HspX/EsxS fusion protein of Mycobacterium tuberculosis along with ISCOMATRIX and PLUSCOM nano-adjuvants after subcutaneous administration in animal model.

Microb Pathog 2021 May 21;154:104842. Epub 2021 Mar 21.

Nanotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address:

Background: Tuberculosis (TB), caused by Mycobacterium tuberculosis (M. tuberculosis), is one of the most common and dangerous infectious diseases in the world. Despite vaccination with BCG, it is still considered as a major health problem. Therefore, design and production of an effective novel vaccine against TB is necessary. Our aim was to evaluate immunogenicity of HspX/EsxS fusion protein of M. tuberculosis along with ISCOMATRIX, PLUSCOM nano-adjuvants and MPLA through the subcutaneous route in mice model.

Methods: HspX/EsxS fused protein of M. tuberculosis was cloned, expressed and purified in the prokaryotic system. ISCOMATRIX and PLUSCOM nano-adjuvants were prepared by film hydration method. Subcutaneous immunization of BALB/c mice was performed by different formulations. IFN-γ, IL-4, IL-17 and TGF-β cytokines levels as well as serum IgG1, IgG2a.

Results: Our results showed that subcutaneous administration of mice with HspX/EsxS along with three adjuvants, ISCOMATRIX, PLUSCOM and MPLA increased immunogenicity of multi-stage fusion protein of M. tuberculosis. Additionally, HspX/EsxS protein + ISCOMATRIX or + PLUSCOM nano-adjuvants induced stronger Th1, IgG2a and IgG1 immune responses compared to MPLA adjuvant. Totally, HspX/EsxS/ISCOMATRIX/MPLA, HspX/EsxS/PLUSCOM/MPLA and two BCG booster groups could significantly induce higher Th1 and IgG2a immune responses.

Conclusion: With regard to ability of ISCOMATRIX, PLUSCOM and MPLA adjuvants to increase immunogenicity of HspX/EsxS protein through induction of IFN-γ and IgG2a immune responses, it seems that these adjuvants and especially ISCOMATRIX and PLUSCOM, could also improve BCG efficacy as a BCG booster.
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http://dx.doi.org/10.1016/j.micpath.2021.104842DOI Listing
May 2021

Response surface methodology optimized electrochemical DNA biosensor based on HAPNPTs/PPY/MWCNTs nanocomposite for detecting Mycobacterium tuberculosis.

Talanta 2021 May 11;226:122099. Epub 2021 Jan 11.

Antimicrobial Resistance Research Center, Department of Medical Bacteriology and Virology, Qaem University Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address:

An important issue in the prognosis of tuberculosis (TB) is a short period between correct diagnosis and start the suitable antibiotic therapy. So, a rapid and valid method for detection of Mycobacterium tuberculosis (M. tb) complex is considered as a necessity. Herein, a rapid, low-cost, and PCR-free DNA biosensor was developed based on multi-walled carbon nanotubes (MWCNTs), polypyrrole (PPy), and hydroxyapatite nanoparticles (HAPNPs) for highly sensitive and specific recognition of M.tb. The biosensor consisted of M.tb ssDNA probe covalently attached to the HANPs/PPy/MWCNTs/GCE surface that hybridized to a complementary target sequence to form a duplex DNA. The M.tb target recognition was based on the oxidation signal of the electroactive Methylene Blue (MB) on the surface of the modified GCE using differential pulse voltammetry (DPV) method. It is worth to mention that for the first time Plackett-Burman (PB) screening design and response surface method (RSM) based on central composite design (CCD) was applied as a powerful and an efficient approach to find optimal conditions for maximum M.tb biosensor performance leading to simplicity and rapidity of operation. The proposed DNA biosensor exhibits a wide detection range from 0.25 to 200.0 nM with a low detection limit of 0.141 nM. The performance of designed biosensor for clinical diagnosis and practical applications was revealed through hybridization between DNA probe-modified GCE and extracted DNA from sputum clinical samples.
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http://dx.doi.org/10.1016/j.talanta.2021.122099DOI Listing
May 2021

pulmonary infection: a case series and literature review.

Respirol Case Rep 2021 Mar 22;9(3):e00719. Epub 2021 Feb 22.

Antimicrobial Resistance Research Center Mashhad University of Medical Sciences Mashhad Iran.

Incidence of pulmonary infection is increasing and diagnosis and treatment are challenging. We surveyed the clinical features, risk factors, diagnosis, and management in 20 patients from northeastern Iran diagnosed by line probe assay and confirmed by sequencing the () region and carried out a literature review using the keywords "pulmonary infection" and "." The mean age of patients was 55.1 years, with 80% female and 90% diagnosed by sputum. Clinical symptoms included severe cough (90%), sputum production (70%), haemoptysis (50%), and chest pain (35%). Comorbidities included a history of tuberculosis (60%), smoking (40%), or chronic obstructive pulmonary disease (20%). Patients were treated with levofloxacin, clarithromycin, and co-trimoxazole. Except for two patients, the clinical symptoms improved. pulmonary infection is increasing in people with underlying diseases. Although choosing the most appropriate treatment remains a challenge, combining successful treatments could be useful in treating these patients.
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http://dx.doi.org/10.1002/rcr2.719DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7898274PMC
March 2021

A case of multidrug-resistant in an elderly woman.

Respirol Case Rep 2021 Mar 1;9(3):e00715. Epub 2021 Feb 1.

Antimicrobial Resistance Research Center Mashhad University of Medical Sciences Mashhad Iran.

is an emerging and spreading pathogen in Iran and little data about its drug susceptibility test (DST) and no standard treatment regimen are available. We report a case of multidrug-resistant . respiratory infection in a 65-year-old woman with a history of previous infection. The patient was treated with clarithromycin, levofloxacin, and cotrimoxazole for one year and eventually died while still suffering from respiratory problems. For DST, broth microdilution method was used according to the Clinical and Laboratory Standards Institute guidelines as well as molecular DST in clinical isolate. was resistant to streptomycin, moxifloxacin, clarithromycin, and cotrimoxazole antibiotics and was sensitive to clofazimine and amikacin antibiotics. Inappropriate use of antibiotics without determining the pattern of antibiotic resistance increases the likelihood of resistance and, for resistant specimens, the need to review the treatment protocol and replace antibiotics. Effectiveness based on antibiotic resistance pattern is essential.
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http://dx.doi.org/10.1002/rcr2.715DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7848708PMC
March 2021

Detection of efflux pump genes in multiresistant Acinetobacter baumannii ST2 in Iran.

Acta Microbiol Immunol Hung 2021 Feb 2. Epub 2021 Feb 2.

11Department of Clinical Psychology, Zahedan University of Medical Sciences, Zahedan, Iran.

Acinetobacter baumannii, as a nosocomial pathogen has become a worldwide concern in recent years. In the current study, the resistance to tetracyclines and colistin were assessed in the isolates from different provinces of Iran.During the timeline of this study, a number of 270 isolates of A. baumannii were collected from tracheal aspirates, wounds, urine and blood cultures. The minimum inhibitory concentration (MIC) for tetracycline, doxycycline, minocycline, tigecycline and colistin were evaluated. Tetracycline resistance genes were assessed by PCR. The mean expression level of adeB, adeJ and adeG were assessed using semi quantitative Real-Time PCR. The clonal relationship of the isolates was evaluated by the repetitive extragenic palindromic PCR (REP-PCR), International Clonal (IC) Lineage Multiplex PCR and multilocus sequence typing (MLST) (Pasteur scheme) methods.The MIC by microdilution method showed that 87.5, 51.4, 28, 0.74 and 0% of the isolates were resistant to tetracycline, doxycycline, minocycline, tigecycline and colistin respectively. The prevalence of tetracycline resistance genes was 99.2, 99.2, 98, 86.7, 10, 3.33, 0.37, 0% for adeB, adeJ, adeG, tetB, tetA(39), tetA, tetM and tetH in tetracycline-resistant isolates. Moreover, the expression level of adeB, adeJ, adeG genes in tigecycline-nonsusceptible A. baumannii (TNAB) strain was higher compared to the tigecycline-susceptible A. baumannii (TSAB). A broad genomic diversity was revealed, but ST2 was the most prevalent ST. Our results indicated that tetracycline resistance in Iran is mediated by resistance-nodulation-cell division (RND) and tetB efflux pumps.
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http://dx.doi.org/10.1556/030.2021.01314DOI Listing
February 2021

Nanotechnology-driven advances in the treatment of diabetic wounds.

Biotechnol Appl Biochem 2020 Oct 12. Epub 2020 Oct 12.

Immunology Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Diabetic foot ulcers (DFUs) are chronic severe complications of diabetes disease and remain a worldwide clinical challenge with social and economic consequences. Diabetic wounds can cause infection, amputation of lower extremities, and even death. Several factors including impaired angiogenesis, vascular insufficiency, and bacterial infections result in a delayed process of wound healing in diabetic patients. Treatment of wound infections using traditional antibiotics has become a critical status. Thus, finding new therapeutic strategies to manage diabetic wounds is urgently needed. Nanotechnology has emerged as an efficient approach for this purpose. This review aimed to summarize recent advances using nanotechnology for the treatment of diabetic wounds.
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http://dx.doi.org/10.1002/bab.2051DOI Listing
October 2020

The overview and perspectives of biosensors and Mycobacterium tuberculosis: A systematic review.

J Cell Physiol 2021 Mar 15;236(3):1730-1750. Epub 2020 Sep 15.

Medical Toxicology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

Tuberculosis (TB) is referred to as a "consumption" or phthisis, which has been a fatal human disease for thousands of years. Mycobacterium tuberculosis (M. tb) might have been responsible for the death of more humans than any other bacterial pathogens. Therefore, the rapid diagnosis of this bacterial infection plays a pivotal role in the timely and appropriate treatment of the patients, as well as the prevention of disease spread. More than 98% of TB cases are reported in developing countries, and due to the lack of well-equipped and specialized diagnostic laboratories, development of effective diagnostic methods based on biosensors is essential for this bacterium. In this review, original articles published in English were retrieved from multiple databases, such as PubMed, Scopus, Google Scholar, Science Direct, and Cochrane Library during January 2010-October 2019. In addition, the reference lists of the articles were also searched. Among 109 electronically searched citations, 42 articles met the inclusion criteria. The highest potential and wide usage of biosensors for the diagnosis of M. tb and its drug resistance belonged to DNA electrochemical biosensors (isoniazid and rifampin strains). Use of biosensors is expanding for the detection of resistant strains of anti-TB antibiotics with high sensitivity and accuracy, while the speed of these sensory methods is considered essential as well. Furthermore, the lowest limit of detection (0.9 fg/ml) from an electrochemical DNA biosensor was based on graphene-modified iron-oxide chitosan hybrid deposited on fluorine tin oxide for the MPT64 antigen target. According to the results, the most common methods used for M. tb detection include acid-fast staining, cultivation, and polymerase chain reaction (PCR). Although molecular techniques (e.g., PCR and real-time PCR) are rapid and sensitive, they require sophisticated laboratory and apparatuses, as well as skilled personnel and expertise in the commentary of the results. Biosensors are fast, valid, and cost-efficient diagnostic method, and the improvement of their quality is of paramount importance in resource-constrained settings.
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http://dx.doi.org/10.1002/jcp.30007DOI Listing
March 2021

A new DNA vaccine expressing HspX-PPE44-EsxV fusion antigens of induced strong immune responses.

Iran J Basic Med Sci 2020 Jul;23(7):909-914

Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

Objectives: Infection with tuberculosis (TB) is regarded as a major health issue. Due to the emergence of antibiotic resistance during TB treatment, prevention via vaccination is one of the most effective ways of controlling the infection. DNA vaccines are developed at a greater pace due to their ability in generating a long-lasting immune response, higher safety compared to the live vaccines, and relatively lower cost of production. In the present study, we evaluated a new DNA vaccine encoding the fusion HspX-PPE44-EsxV antigens, separately, and in combination with Bacillus Calmette-Guérin (BCG) administration, in a prime-boost method in mice.

Materials And Methods: A novel DNA vaccine encoding HspX-PPE44-EsxV fusion antigen of was constructed, and RT-PCR and Western blot analysis were performed to verify the expression of the antigen. Female BALB/c mice were divided into five groups (PBS, BCG, pcDNA3.1 (+) vector, pDNA/HspX-PPE44-EsxV vaccine, and the BCG-prime boost groups). In order to evaluate the immunogenicity of the recombinant vector, BALB/c mice were injected with 100 μg of pDNA at 2-week intervals. Then, cytokine assay was conducted using eBioscience ELISA kits (Ebioscience, AUT) according to manufacturers' instructions to evaluate the concentrations of IL-4, IL-12, TGF-β, and IFN-γ.

Results: The concentrations of INF-γ, IL-12, and TGF-beta were significantly increased compared to the control groups (<0.001). INF-γ and IL-12 production were increased significantly in pDNA/HspX-PPE44-EsxV+BCG group compared to pDNA/HspX-PPE44-EsxV group (<0.001).

Conclusion: This study showed that the present DNA vaccine could induce a high level of specific cytokines in mice. It was also shown that using this DNA vaccine in a BCG prime-boost protocol can produce significant amounts of IFN-γ, IL-12, and TGF-β.
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http://dx.doi.org/10.22038/ijbms.2020.38521.9171DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7395183PMC
July 2020

A novel formulation of Mtb72F DNA vaccine for immunization against tuberculosis.

Iran J Basic Med Sci 2020 Jun;23(6):826-832

Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

Objectives: (), an intracellular pathogen, causes 1.5 million deaths globally. Bacilli Calmette-Guérin (BCG) is commonly administered to protect people against infection; however, there are some obstacles with this first-generation vaccine. DNA vaccines, the third generation vaccines, can induce cellular immune responses for tuberculosis (TB) protection. In this study, optimized DNA vaccine (pcDNA3.1-Mtb72F) entrapped in poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) was used to achieve higher immunogenicity.

Materials And Methods: Plasmid Mtb72F was formulated in PLGA NPs using double emulsion method in the presence of TB10.4 and/or CpG as an adjuvant. Female BALB/c mice were immunized either with NP-encapsulated Mtb72F or naked Mtb72F with or without each adjuvant, using the BCG-prime DNA boost regimen.

Results: These NPs were approximately 250 nm in diameter and the nucleic acid and protein encapsulation efficiency were 80% and 25%, respectively. The NPs smaller than 200 nm are able to promote cellular rather than humoral responses. The immunization with the formulation consisting of Mtb72F DNA vaccine and TB10.4 entrapped in PLGA NPs showed significant immunogenicity and induced predominantly interferon-ɣ (IFN-ɣ) production and higher INF-ɣ/interleukin-4 (IL-4) ratio in the cultured spleen cells supernatant.

Conclusion: PLGA NPs loaded with Mtb72F DNA-based vaccine with TB10.4 could be considered as a promising candidate for vaccination against TB. These results represent an excellent initial step toward development of novel vaccine for TB protection.
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http://dx.doi.org/10.22038/ijbms.2020.41806.9881DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7351443PMC
June 2020

Development of a Cost-Effective Line Probe Assay for Rapid Detection and Differentiation of Species: A Pilot Study.

Rep Biochem Mol Biol 2020 Jan;8(4):383-393

Immunobiochemistry Laboratory, Immunology Research Center, Medical School, Mashhad University of Medical Sciences, Mashhad, Iran.

Background: The line probe assay (LPA) is one of the most accurate diagnostic tools for detection of different species. Several commercial kits based on the LPA for detection of Mycobacterium species are currently available. Because of their high cost, especially for underdeveloped and developing countries, and the discrepancy of non-tuberculous mycobacteria (NTM) prevalence across geographic regions, it would be reasonable to consider the development of an in-house LPA. The aim of this study was to develop an LPA to detect and differentiate mycobacterial species and to evaluate the usefulness of PCR-LPA for direct application on clinical samples.

Methods: One pair of biotinylated primers and 15 designed DNA oligonucleotide probes were used based on multiple aligned internal transcribed spacer (ITS) sequences. Specific binding of the PCR-amplified products to the probes immobilized on nitrocellulose membrane strips was evaluated by the hybridization method. Experiments were performed three times on separate days to evaluate the assay's repeatability. The PCR-LPA was evaluated directly on nine clinical samples and their cultivated isolates.

Results: All 15 probes used in this study hybridized specifically to ITS sequences of the corresponding standard species. Results were reproducible for all the strains on different days. Mycobacterium species of the nine clinical specimens and their cultivated isolates were correctly identified by PCR-LPA and confirmed by sequencing.

Conclusion: In this study, we describe a PCR-LPA that is readily applicable in the clinical laboratory. The assay is fast, cost-effective, highly specific, and requires no radioactive materials.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7275829PMC
January 2020

Rapid and label-free electrochemical DNA biosensor based on a facile one-step electrochemical synthesis of rGO-PPy-(L-Cys)-AuNPs nanocomposite for the HTLV-1 oligonucleotide detection.

Biotechnol Appl Biochem 2020 Jun 15. Epub 2020 Jun 15.

Inflammation and Inflammatory Disease Research Centre, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Human T cell leukemia virus type 1 (HTLV-1) as the first human retrovirus is currently a serious endemic health challenge. Despite the use of assorted molecular or serological assays for HTLV-1 detection, there are several limitations due to the lack of a confirmatory test that may affect the accuracy of the results. Herein, a novel label-free biosensor for the detection of HTLV-1 Tax gene has been reported. An electrochemical facile ecofriendly synthesis method has been demonstrated based on a synthesis of nanocomposite of reduced graphene oxide, polypyrrole, and gold nanoparticles (rGO-PPy-(l-Cys)-AuNPs) deposited on the surface of screen-printed carbon electrode. Electrochemical techniques were used to characterize and study the electrochemical behavior of the rGO-PPy-(l-Cys)-AuNPs, which exhibited a stable reference peak at 0.21 V associated with hybridization forms by applying the differential pulse voltammetry. The designed DNA biosensor presented a wide linear range from 0.1 fM to 100 µM and a low detection limit of 20 atto-molar. The proposed biosensor presented in this study provides outstanding selectivity, sensitivity, repeatability, and reproducibility.
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http://dx.doi.org/10.1002/bab.1973DOI Listing
June 2020

Genotypic and phenotypic characterization of Mycobacterium tuberculosis resistance against fluoroquinolones in the northeast of Iran.

BMC Infect Dis 2020 Jun 1;20(1):390. Epub 2020 Jun 1.

Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

Background: Fluoroquinolones are broad-spectrum antibiotics that are recommended, and increasingly important, for the treatment of multidrug-resistant tuberculosis (MDR-TB). Resistance to fluoroquinolones is caused by mutations in the Quinolone Resistance Determining Region (QRDR) of gyrA and gyrB genes of Mycobacterium tuberculosis. In this study, we characterized the phenotypic and genotypic resistance to fluoroquinolones for the first time in northeast Iran.

Methods: A total of 123 Mycobacterium tuberculosis isolates, including 111 clinical and 12 collected multidrug-resistant isolates were studied. Also, 19 WHO quality control strains were included in the study. The phenotypic susceptibility was determined by the proportion method on Löwenstein-Jensen medium. The molecular cause of resistance to the fluoroquinolone drugs ofloxacin and levofloxacin was investigated by sequencing of the QRDR region of the gyrA and gyrB genes.

Results: Among 123 isolates, six (4.8%) were fluoroquinolone-resistant according to phenotypic methods, and genotypically three of them had a mutation at codon 94 of the gyrA gene (Asp→ Gly) which was earlier reported to cause resistance. All three remaining phenotypically resistant isolates had a nucleotide change in codon 95. No mutations were found in the gyrB gene. Five of the 19 WHO quality control strains, were phenotypically fluoroquinolone-resistant, four of them were genotypically resistant with mutations at codon 90, 91 of the gyrA gene and one resistant strain had no detected mutation.

Conclusions: Mutation at codon 94 of the gyrA gene, was the main cause of fluoroquinolone resistance among M. tuberculosis isolates in our region. In 3/6 fluoroquinolone-resistant isolates, no mutations were found in either gyrA or gyrB. Therefore, it can be concluded that various other factors may lead to fluoroquinolone resistance, such as active efflux pumps, decreased cell wall permeability, and drug inactivation.
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http://dx.doi.org/10.1186/s12879-020-05112-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7268510PMC
June 2020

The effects of high doses of vitamin D on the composition of the gut microbiome of adolescent girls.

Clin Nutr ESPEN 2020 02 5;35:103-108. Epub 2019 Dec 5.

Metabolic Syndrome Research Center, School of Medicine, Mashhad University of Medical Science, Mashhad, Iran. Electronic address:

Background: Animal studies suggest that vitamin D can change the gut microbiome. The primary aim of this study was to evaluate the effect of a high dose supplementation of vitamin D on the composition of the gut microbiome.

Methods: After DNA extraction, TaqMan assays were used for the quantitation of selected microbiome in the feces of 50 adolescent girls before and after vitamin D supplementation.

Results: The expression fold changes for Enterococcus, Bifidobacterium, Lactobacillus, Bacteroidetes and Firmicutes were; 1.05, 1.20, 0.76, 0.28 and 1.50 respectively. Bacteroidetes and Lactobacillus fell by 72% (P < 0.0001) and 24% (P = 0.006) respectively, whilst Firmicutes and Bifidobacterium were increased by 1.5 (P < 0.0001), 1.2 (P < 0.0001) fold after supplementation.

Conclusion: Our results suggested that a high dose supplementation of vitamin D alter the human gut microbiome composition. Future studies are required for a better understanding of the mechanisms by which vitamin D affects the gut microbiome.
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http://dx.doi.org/10.1016/j.clnesp.2019.10.020DOI Listing
February 2020

The roles of latency-associated antigens in tuberculosis vaccines.

Indian J Tuberc 2019 Oct 1;66(4):487-491. Epub 2019 May 1.

Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address:

Tuberculosis (TB) is a re-emerging disease and is caused by Mycobacterium tuberculosis (M. tuberculosis). TB is currently one of the leading causes of morbidity and mortality, worldwide. The only available vaccine against TB infection, Bacillus Calmette-Guérin (BCG), fails to adequately protect against reactivation of latent infections in adults. Furthermore, recently developed subunit vaccines, which are in various stages of clinical trials, are all prophylactic vaccines based on proteins expressed in replicating stage of M. tuberculosis and they are not preventive of reactivation of latent TB infection. Thus, an appropriate subunit post-exposure vaccine needs to be developed to control all forms of TB infection. To produce a multi-stage subunit vaccine, scientists should combine the early secreted M. tuberculosis antigens with latency antigens. For this purpose, some latency proteins are known which could be important antigens in the production of specific humoral and cellular immune responses in latent M. tuberculosis infected individuals. Several studies have evaluated the immunogenicity of these proteins in improving the TB vaccines. The present study is a comprehensive review of several studies on the role of the latency antigens in the development of TB vaccines. Overall, the studies indicate that the latency-associated antigens including the resuscitation-promoting factors, the Dormancy of survival regulon (DosR) proteins and the starvation stimulant proteins are potential candidates for the development of subunit vaccines against TB infection.
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http://dx.doi.org/10.1016/j.ijtb.2019.04.012DOI Listing
October 2019

Prevalence of CCR5delta32 in Northeastern Iran.

BMC Med Genet 2019 11 15;20(1):184. Epub 2019 Nov 15.

Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, P.O Box: 9196773117, Mashhad, IR, Iran.

Background: A 32-base pair deletion (∆32) in the open reading frame (ORF) of C-C motif chemokine receptor 5 (CCR5) seems to be a protective variant against immune system diseases, especially human immunodeficiency virus type 1 (HIV-1). We aimed to assess the frequency of CCR5∆32 in the healthy Iranian population.

Methods: In this study, 400 normal samples from Khorasan, northeastern Iran, were randomly selected. The frequency of CCR5∆32 carriers was investigated using PCR analysis. Allele prevalence and the fit to the Hardy-Weinberg equilibrium were analyzed.

Results: The prevalence of CCR5∆32 in the northeastern population of Iran was 0.016. Four hundred samples were studied, among which one with CCR5 and 11 with CCR5 genotype were detected.

Conclusion: This study was the first investigation for an assessment of the prevalence of CCR5∆32 in northeastern Iran. The low prevalence of CCR5∆32 allele in the Iranian population may result in the increased susceptibility to HIV-1. In addition, this prevalence is the same as that of reported in East Asia, while is lower than that in the Europeans.
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http://dx.doi.org/10.1186/s12881-019-0913-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6858674PMC
November 2019

Design and Construction of a Eukaryotic Cloning Vector Encoding the mpt51 Gene of .

Rep Biochem Mol Biol 2019 Apr;8(1):32-35

Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

Background: Tuberculosis (TB) is the leading cause of death by infectious diseases worldwide, and especially prevalent in developing countries. Several vaccines against TB have been developed, recently. The aim of the present study was to design and construct a cloning vector encoding gene.

Methods: DNA was extracted from MTB H37Rv strain. Gene-specific primers were designed using Gene Runner software and the gene was amplified by PCR. The amplified fragment and pcDNA3.1(+) cloning vector were both digested with restriction enzymes, the fragment was ligated into the vector, and the TOP10 strain were transformed by the recombinant plasmid. Positive clones were identified by colony PCR, restriction enzyme digestion, and DNA sequencing.

Results: The gene was successfully cloned into pcDNA3.1(+). A 6400 bp band for the pcDNA3.1(+)/ recombinant plasmid and a 926 bp band for were observed by colony PCR, and restriction enzyme digestion on agarose gels. The DNA sequence was 100% homologous with the fragment of H37Rv in GenBank.

Conclusion: In the current study, the gene of MTB was correctly cloned into pcDNA3.1(+). The expression of this recombinant vector can be studied in eukaryotic cells. Moreover, it is possible to determine the efficacy of this vector as a DNA vaccine candidate, and to test its protective function compared to BCG in animal models in future.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6590942PMC
April 2019

New anthropometric indices in the definition of metabolic syndrome in pediatrics.

Diabetes Metab Syndr 2019 May - Jun;13(3):1779-1784. Epub 2019 Mar 19.

Metabolic Syndrome Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address:

Pediatrics metabolic syndrome (MetS) may be associated with the risk of development of chronic diseases in adulthood; however, the definition of pediatric MetS is unclear, and may vary with ethnicity. The primary goal of this study was to determine the best anthropometric predictors for pediatric MetS. For this purpose, 988 high school girls were recruited. Anthropometric indices and biochemical parameters were measured using standard procedures. The adapted MetS for pediatrics, including the IDF, NCEP, and two modified-NCEPs (Cook's and DeFerranti's) were used to establish a diagnosis of MetS. Statistical analysis was performed using SPSS and MedCalc softwares. Except for body frame size (r), the values for anthropometric indices were significantly lower in an individual without MetS. Waist to height (WHtR), BMI and hip circumference (HiC) showed the strongest association with the different MetS definitions. For the IDF definition, the highest sensitivity and specificity were observed for HiC (100.0, 85.2) and WHtR (100.0, 84.7); while for the NCEP definition, the r index showed the highest sensitivity (85.0); but low specificity made it inapplicable. For the Cook's definition of MetS, wrist circumference (WrC), HiC, WHtR, BMI and SR had similar sensitivity values with WC (92.9%), and HiC (85.3%) have the highest specificity. WHtR (86.05, 80.5), SR (86.05, 82.7) and HiC (76.7, 87.0) sensitivity and specificity were the best indexes for DeFerranti's criteria. Based on this date, we concluded that HiC and WHtR might be helpful as auxiliary indexes for pediatric MetS definition; however, further studies are required in both genders.
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http://dx.doi.org/10.1016/j.dsx.2019.03.032DOI Listing
December 2019

A method for improving the efficiency of DNA extraction from clotted blood samples.

J Clin Lab Anal 2019 Jul 10;33(6):e22892. Epub 2019 May 10.

Biochemistry of Nutrition Research Center, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Background: The efficient and rapid extraction of high-quality genomic DNA from clotted blood samples, which normally have a low yield and poor quality, is an important factor in genomic research. The objective of this study was to develop a simple and safe technique for dispersing the blood clots by the ball bearing metal shots. Normally, such clot samples may not have an acceptable yield by conventional DNA extraction methods. Also, in the present study, we have further investigated to improve salting-out DNA extraction methods.

Methods: Initially, 500 µL phosphate-buffered saline (PBS) (1×) and two ball bearing metal shots were added to each tube of the clotted blood sample and then were gently rotated in an electric laboratory rotator for 1 hour at room temperature (18-25°C). Genomic DNA was then extracted from samples using a modified salting-out method and a modified QIAamp DNA Blood Midi Kit and was compared with QIAamp DNA Blood Midi Kit as a control. An assessment of the concentration and quality of the extracted DNA was performed using the UV-visible spectrophotometer. The isolated DNA proved amenable to PCR amplification and gel electrophoresis.

Results: The yield and purity of DNA obtained by these three methods were significantly different (P < 0.001), with a higher yield in the modified salting-out method.

Conclusions: Our proposed modified salting-out method is simple and efficient for the isolation of DNA from old blood clot samples. It is both easy to use and is of low cost in routine laboratory tasks.
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http://dx.doi.org/10.1002/jcla.22892DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6642314PMC
July 2019

Designing and Construction of a Cloning Vector Containing Gene of .

Tanaffos 2018 Mar;17(3):198-202

Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

Background: Tuberculosis caused by , remains as one of the leading causes of deaths worldwide, with nearly two million death cases annually. BCG (Bacille Calmette-Guerin) continues to be the most widely used vaccine in the world, but the protective immunity differs in different parts of the world. Accordingly, new strategies including DNA vaccines are essentially needed. This study was aimed to design and construct a cloning vector containing gene of .

Materials And Methods: H37Rv was cultured on Lowenstein Jensen medium, and genomic DNA was extracted. The gene was amplified by PCR using designed specific primers. After the digestion of and pcDNA3.1 (+) by HI and RI restriction enzymes, the fragment was ligated into the digested vector using enzyme. Then, the recombinant vector was transformed into competent ( TOP10 strain. To confirm the colonies of transformed bacteria, antibiotic resistance, colony-PCR, restriction enzyme digestion and DNA sequencing were used.

Results: To confirm the clones, colony-PCR using specific primers was performed and the fragment of 718 bp was observed by gel electrophoresis. Clones were also verified by restriction enzyme digestion using HI and RI restriction enzymes and the 718 bp fragment was observed. Furthermore, results of DNA sequencing showed 100% homology with the fragment of H37Rv in GenBank.

Conclusion: In this study, the fragment was successfully cloned in pcDNA3.1 (+) vector. This construct can be used in future studies as a DNA vaccine in animal models to induce immune system responses.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6428376PMC
March 2018

ISAba1/bla family is the predominant cause of carbapenem resistance in Acinetobacter baumannii and Acinetobacter nosocomialis in Iran.

Infect Genet Evol 2019 07 19;71:60-66. Epub 2019 Mar 19.

Liver and Digestive Research Center, Kurdistan University of Medical Sciences, Sanandaj, Iran; Department of Microbiology, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran. Electronic address:

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http://dx.doi.org/10.1016/j.meegid.2019.03.009DOI Listing
July 2019

Investigation of the Mutations Causing Rifampin Resistance by Rapid Screening in in North-East of Iran.

Iran J Pathol 2018 25;13(4):429-437. Epub 2018 Sep 25.

Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

Background And Objectives: The incidence of rifampin-resistant strains of has attracted more attention than the tuberculosis infection due to laborious treatment and control. Recognizing the genotypes involving in drug resistance via multiplex PCR, a simple and rapid genotyping method, is an emergency for better treatment and control of tuberculosis. This study was designed to specify the frequency of rifampin-resistant strains of isolated from patients by multiplex allele-specific Polymerase Chain Reaction assay (MAS-PCR).

Methods: In this study, 88 positive samples were included from Qaem Hospital, Mashhad. MAS-PCR was used to detect the rifampin resistance associated mutations in gene.

Results: Mutations in three codons of gene causing rifampin resistance were detected in 51 isolates (58.96%). The detected mutations in codons 531, 526, and 516 were 55.68%, 38.63%, and 13.63%, respectively. The simultaneous mutations were detected in 11 isolates (12.50%) in codons 531, 526 and 516, in 21 isolates (23.86%) in codons 531 and 526, and in one isolate (1.13%) in codons 526 and 516.

Conclusion: According to the results of this study, the frequency of rifampin-resistant strains of isolated from Khorasan province patients (North-East of Iran) was high. The developed MAS-PCR assay can be used for rapid detection in clinical diagnostic laboratories in areas with high prevalence of multidrug-resistant strains. In this respect, MAS-PCR is simple, rapid, and highly sensitive method for drug susceptibility tests for detecting multidrug-resistant .
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6358560PMC
September 2018

Designing and Construction of a Cloning Vector Encoding Fragments of as a DNA Vaccine Candidate.

Iran J Pathol 2018 25;13(4):403-407. Epub 2018 Sep 25.

Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

Background & Objective: Tuberculosis (TB) remains a major cause of death around the world. Bacillus Calmette Guérin (BCG) is the only vaccine used in TB prevention that has a protective effect in children, but its effectiveness declines in adults. Design and development of new vaccines is the most effective way against TB.The aim of this study was to design and construct a DNA vaccine encoding and fusion genes of .

Methods: First, fragment was amplified by PCR method. The pcDNA3.1+/ plasmid was transformed into JM109 and then extracted. The gene and pcDNA3.1+/ plasmid were both digested with RI and HI restriction enzymes followed by ligation of 51 fragment into the digested vector. The recombinant plasmid containing and was subsequently transformed into competent TOP10 strain. The clones were confirmed by colony-PCR, restriction enzyme digestion and sequencing.

Results: Using agarose gel electrophoresis, a 926 bp fragment corresponded to was observed. Digestion of the vector pcDNa3.1+/ and gene was confirmed by electrophoresis. Then, the pcDNA3.1+/ plasmid was extracted. Sequencing results confirmed the accuracy of the desired plasmid.

Conclusion: In this study, we constructed a cloning vector encoding gene of . The eukaryotic expression of this vector can be confirmed in future studies. It can be considered as a DNA vaccine in animal models later. Successful cloning provides a basis for the development of new DNA vaccines against TB.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6358561PMC
September 2018

Association Between Trace Element Status and Depression in HTLV-1-Infected Patients: a Retrospective Cohort Study.

Biol Trace Elem Res 2019 Sep 4;191(1):75-80. Epub 2019 Feb 4.

Metabolic Syndrome Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Depression and Anxiety are two important public health problems that are known to be associated with viral infections. The association between the intake of nutrients such as zinc and copper with symptoms of depression has been studied previously. The aim of the current study was to investigate the association between depression with human T cell lymphotropic virus type 1 (HTLV-1) infection and serum content of zinc and copper in a large Iranian population cohort. The study population consisted of 279 HTLV-1-positive patients who were identified after recruitment as part of a large cohort study: the Mashhad Stroke and Heart Association Disorder (MASHAD) study. They were divided into two groups of diagnosed with or without depression based on their symptoms. Serum zinc and copper levels of all subjects were measured using the flame atomic absorption spectrometry. The population sample comprised of 279 individuals infected with HTLV-1 of whom 192 (68.8%) were women. The mean serum zinc in the group with and without depression was 78.69 ± 13.79 μg/dl and 86.87 ± 19.44 μg/dl, respectively (p < 0.001). Also, the serum copper level was higher in the depressive group (116.75 ± 39.56) than in the non-depressive group (104.76 ± 30.77) (p 0.004). The association between serum zinc and copper with depression in HTLV-1-infected patients which was shown in this study could be considered in the treatment strategies in these patients.
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http://dx.doi.org/10.1007/s12011-018-1613-6DOI Listing
September 2019

Development of Preventive Vaccines for Hepatitis C Virus E1/E2 Protein.

Iran J Pathol 2018 17;13(2):113-124. Epub 2018 Jul 17.

Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

Hepatitis C virus (HCV) is responsible for a vast majority of liver failure cases. HCV is a kind of blood disease estimated to chronically infect 3% of the worlds population, causing significant morbidity and mortality. Therefore, a complete knowledge of humoral responses against HCV, resulting antibodies, and virus-receptor and virus-antibody interactions, are essential to design a vaccine. HCV epitopes or full sequence of HCV proteins can induce HCV specific immune responses. In fact, structural proteins are usually the main target of humoral responses and non-structural proteins are usually the main target of cellular responses. Hence, various vaccines based on distinct antigenic combinations are developed to prevent HCV infection and the current study tried to summarize them.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6339490PMC
July 2018

Human T lymphotropic virus type 1 and risk of cardiovascular disease: High-density lipoprotein dysfunction versus serum HDL-C concentrations.

Biofactors 2019 May 29;45(3):374-380. Epub 2019 Jan 29.

Metabolic Syndrome Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

High-density lipoprotein (HDL) is thought to be protective against cardiovascular disease (CVD), and HDL dysfunction is considered to be a risk factor for CVD. It is unclear whether there is an association between Human T lymphotropic virus type 1 (HTLV1) infection and CVD risk. We have assessed HDL lipid peroxidation (HDLox) as a marker of HDL dysfunction and CVD risk in a subgroup of the MASHAD cohort study. One hundred and sixty two individuals including 50 subjects positive for HTLV1 infection and 112 individuals negative for HTLV1 infection were recruited. Anthropometric and biochemical parameters including serum hs-CRP, fasted lipid profile (HDL-C, LDL, triglycerides, and cholesterol), and fasting blood glucose were determined. Serum HDLox was also measured in the study participants. Multivariate analyses were used to evaluate the association between serum HDLox and HTLV1 infection. None of the traditional CVD risk factors were associated with HTLV1 infection, including serum HDL-C. However, serum HDLox was independently associated with the presence of HTLV1 infection. Logistic regression analysis showed that subjects who were positive for HTLV1 infection were also significantly more likely than uninfected individuals to have higher HDLox (odds ratio 9.35, 95%CI: 3.5-24.7; P < 0.001). HDLox was increased approximately 20% (P < 0.001) in infected subjects compared to the uninfected group. Serum HDLox is a marker of CVD risk factor and increased in individuals affected by HTLV1 infection compared to healthy subjects. © 2019 BioFactors, 45(3):374-380, 2019.
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http://dx.doi.org/10.1002/biof.1489DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6548577PMC
May 2019

Association between serum zinc and copper levels and antioxidant defense in subjects infected with human T-lymphotropic virus type 1.

J Blood Med 2019 27;10:29-35. Epub 2018 Dec 27.

Metabolic Syndrome Research Center, Mashhad University of Medical Sciences, Mashhad, Iran,

Introduction: Copper (Cu) and zinc (Zn) are important trace elements that are also structural ions of superoxide dismutase (SOD), which reduce oxidative stress. Zinc deficiency and excess copper have been reported to be associated with inflammation. The human T-lymphotropic virus type 1 (HTLV-1) is a retrovirus, which is believed to cause systemic inflammation. The aim of this study is to measure levels of Zn, Cu, SOD, and prooxidant-antioxidant balance (PAB) in HTLV-1-positive patients and investigate the association between serum Zn and Cu concentrations and levels of oxidative stress in them.

Methods: The serum samples of 1,116 subjects who had participated in the "Mashhad Stroke and Heart Atherosclerotic Disorder" study, including 279 HTLV-1-positive and 837 HTLV-1-negative patients, were used. Levels of Zn, Cu, SOD, and PAB were measured.

Results: Zinc and SOD levels were lower in the HTLV-1-positive group; however, the difference was statistically significant only for the level of SOD (=0.003). On the other hand, levels of copper and PAB were significantly higher in HTLV-1 positive subjects; =0.004 and =0.002, respectively.

Conclusion: In HTLV-infected patients, serum Zn concentration is lower and Cu concentration is higher than healthy controls. This altered situation might be either primary or secondary to HTLV-1 infection, which should be investigated in larger studies. We showed that SOD is significantly lower in HTLV-1-infected subjects. As in some other viruses that evolve different mechanisms to potentiate virus replication by changing the physiologic condition of host cells, HTLV-1 too probably decreases the activity of copper-zinc SOD1 by suppressing its gene.
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http://dx.doi.org/10.2147/JBM.S184913DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6312056PMC
December 2018

Current approaches for detection of human T-lymphotropic virus Type 1: A systematic review.

J Cell Physiol 2019 08 11;234(8):12433-12441. Epub 2019 Jan 11.

Division of Medical Education, Brighton & Sussex Medical School, Falmer, Brighton, Sussex, UK.

Background: Human T-lymphotropic virus Type 1 (HTLV-1) is a retrovirus that is endemic in some regions of the world. It is known to cause several diseases like adult T-cell leukemia (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Serology and molecular methods have been used to detect this virus. Of these, enzyme-linked immunosorbent assay (ELISA) is used as a primary screening method and this is usually followed by western blotting (WB) and polymerase chain reaction (PCR) methods as confirmatory tests. We conducted a systematic review of the different techniques used in the diagnosis of HTLV-1 infection.

Materials And Methods: Our search was limited to original papers in the English language from 2010 to 2018 using several databases including Pubmed, Scopus, Google Scholar, Iranmedex, and Scientific Information Database. A manual search of references provided in the included papers was also performed.

Results: Of 101 electronically searched citations, 43 met the inclusion criteria. ELISA is commonly used for qualitative and screening detection, and WB and PCR techniques are used to confirm infection.

Conclusion: Among all the reported methods for detection of HTLV-1, only serological and molecular tests are used as the most common technical assays for HTLV-1. The ELISA assay, without a confirmatory test, has several limitations and affect the accuracy of the results. Owing to the prevalence of HTLV-1 and limitations of the current detection methods, further evaluation of the accuracy of these methods is needed. There are new opportunities for applying novel technological advances in microfluidics, biosensors, and lab-on-a-chip systems to perform HTLV-1 diagnostics.
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http://dx.doi.org/10.1002/jcp.28087DOI Listing
August 2019

Shedding light on the EpCAM: An overview.

J Cell Physiol 2019 08 9;234(8):12569-12580. Epub 2019 Jan 9.

Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran.

The epithelial cell adhesion molecule (EpCAM) is a Type I transmembrane superficial glycoprotein antigen that is expressed on the surface of basolateral membrane of multiple epithelial cells with some exceptions such as epidermal keratinocytes, hepatocytes, thymic cortical epithelial cells, squamous stratified epithelial cells, and myoepithelial cells that do not express the molecule. The molecule plays a pivotal role in the structural integrity, adhesion of the epithelial tissues and their interaction with the underlying layers. EpCAM prevents claudin-7 and claudin-1 molecules from degradation, thereby, decreasing the number of tight junctions and cellular interconnections, and promoting the cells toward carcinogenic transformation. Moreover, the mutations in the EpCAM gene lead to congenital tufting enteropathy, severe intestinal epithelium homeostasis disorders, and Lynch and Lynch syndrome. Overexpression of EpCAM on stem cells of some cancers and the presence of this molecule on circulating tumor cells (CTCs) makes it a promising candidate for cancer diagnosis as well as tracing and isolation of CTCs.
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http://dx.doi.org/10.1002/jcp.28132DOI Listing
August 2019