Publications by authors named "Mojtaba Sankian"

137 Publications

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

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

Protective immune response against P32 oncogenic peptide-pulsed PBMCs in mouse models of breast cancer.

Int Immunopharmacol 2021 Apr 9;93:107414. Epub 2021 Feb 9.

Immunology Research Center, Inflammation and Inflammatory Diseases Division, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address:

High expression of p32 in certain tumors makes it a potential target for immunotherapy. In the present study, the first goal was to design multi-epitope peptides from the P32 protein and the second goal was to compare the prophylactic effects of DCs- and PBMCs- based vaccines by pulsing them with designed peptides. For these purposes, 160 BALB/c mice were vaccinated in 5 different subgroups of each 4 peptides using PBS (F1-4a), F peptides alone (F1-4b), F peptides with CpG-ODN (F1-4c), F peptides with CpGODN and DCs (F1-4d), and F peptides with CpG-ODN and PBMCs (F1-4e). We found a significantly higher interferon-γ (IFN-γ) and granzyme B levels in T cells of F4d and F4e subgroups compared to control (p ≤ 0.05). The result of challenging spleen PBMCs of vaccinated mice with 4T1 cells showed significant up- and down- regulation of Fas ligand (FasL) and forkhead box P3 (Foxp3) gene expression between F4d and F4e subgroups with control, respectively. In addition, a significant change was seen in Caspase3 gene expression of F4d subgroup compared to control (p ≤ 0.05). Supernatant levels of IFN-γ and perforin were significantly increased in F4d and F4e subgroups compared to control. Consequently, significantly lower tumor sizes and prolonged survival time were detected in F4d and F4e subgroups compared to control after challenging mice with 4T1 cells. Accordingly, these results demonstrated that PBMCs pulsed F4 peptide-based vaccine could induce a protective immune response while it is a simple and less expensive vaccine.
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http://dx.doi.org/10.1016/j.intimp.2021.107414DOI Listing
April 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

Preventive Cancer Vaccination by P5 HER-2/neo Derived Peptide-pulsed Peripheral Blood Mononuclear Cells in Mouse Model of Breast Cancer.

Biochem Cell Biol 2021 Jan 4. Epub 2021 Jan 4.

Mashhad University of Medical Sciences, 37552, Mashhad, Iran (the Islamic Republic of);

This study aimed to compare the prophylactic effects of dendritic cells (DCs) and peripheral blood mononuclear cells (PBMCs) based vaccines by pulsing them in vitro with p5 peptide. The different groups of mice were injected by free peptide or peptide pulsed with DCs or PBMCs. Two weeks after the last boosting dose, immunological tests were performed on splenocyte suspensions of three mice, and the remaining mice in each group were evaluated for tumor growth and survival. The IFN-γ, granzyme B, and IL-10 were detected in T cells. Additionally, IFN-γ and perforin as well as mRNA levels of some genes associated with immune responses were assessed after challenging of splenocytes with TUBO cells. A significant increase was observed in frequency of CD4+ IFN-γ+, CD8+ IFN-γ+ and CD8+ granzymeB+ T cells, and the perforin of supernatants in DC and PBMC groups. A significant Fas ligand (FasL) and forkhead box P3 (Foxp3) expression was observed in DC and PBMC groups. These responses led to lower tumor sizes and longer survival time in tumor mice model. The efficacy of this PBMC-based vaccine in improving the protective immune response makes it a simpler and less expensive candidate vaccine compared to DCs-based vaccines.
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http://dx.doi.org/10.1139/bcb-2020-0559DOI Listing
January 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

Diabetes up-regulated collagen IV and laminin α5 genes in mRNA and protein levels in seminiferous tubules of C57BL/6 adult mice.

Cell Mol Biol (Noisy-le-grand) 2020 Jul 31;66(5):162-168. Epub 2020 Jul 31.

Department of Anatomy and Cell Biology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Diabetes is a disease associated with impairment of the male reproductive system that causes complications such as decreased testosterone, the diameter of the seminiferous tubule, libido, and fertility. Extracellular matrix (ECM) molecules are involved in testicular development and spermatogenesis. Laminin and collagen are key proteins in seminiferous tubule basement membrane and play an important role in spermatogenesis. The present study was conducted to investigate the effect of diabetes on collagen IV and laminin α5 changes in mice testis. In this experimental study, 40 mice (C57BL/6) were divided randomly into 4 groups: 1) Control group: without intervention, 2) Diabetic group: treated mice with 50 mg/kg streptozotocin (STZ), 3) Diabetic + Insulin group: treated mice with STZ and insulin, and 4) Sham group: received citrate buffer. After 35 days, the left testes of all specimens were used for Real-Time PCR while their right testes were applied for immunohistochemical study and Periodic acid-Schiff (PAS) staining. This study showed that gene expression and immunoreactivity of laminin α5 and collagen IV were significantly increased in diabetic mice compared to other groups (P<0.05). Also, PAS staining showed the thickness of seminiferous tubule basement membrane in the Diabetic group compared to other group increased significantly (p<0.05). In Diabetic + Insulin compared to Diabetic group, gene expression, the intensity of immunoreactivity and thickness of seminiferous tubule basement membrane decreased significantly (P<0.05). Our findings indicated that diabetes causes up-regulation of collagen IV and laminin α5 in mRNA and protein levels in the seminiferous tubule basement membrane and may cause disorder in spermatogenesis in mice.
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July 2020

Oral Immunotherapy Using Probiotic Ice Cream Containing Recombinant Food-Grade Which Inhibited Allergic Responses in a BALB/c Mouse Model.

J Immunol Res 2020 24;2020:2635230. Epub 2020 Sep 24.

Department of Food Science and Technology, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.

This study was conducted to evaluate the effects of recombinant probiotic bacteria as a candidate for oral vaccine with the potential of treating allergy to pollens. The main gene of this allergen, Ama r 2, was cloned into the food grade plasmid pNZ7025 and then was electrotransformed into the food grade NZ1330. No expression was observed in the primary structure due to the distance between the ribosome binding site and the start codon. Therefore, the vector structure was corrected using the site-directed mutagenesis (SDM) technique. The cell extract of this strain was used for assessing the expression of the recombinant allergen in western blot analysis, and the existence of this protein with a molecular weight of 14.2 kDa was confirmed. To evaluate the efficacy of this strain in the treatment of allergies as an oral vaccine, probiotic ice cream was prepared. After the sensitization of mice, the treatment was performed by oral immunotherapy for 4 weeks, 4 to 5 times per week. 20 l of functional ice cream with 10 CFU/ml of r- NZ1330 significantly reduced the serum IgE level. The levels of IFN- and TGF- cytokines increased in the 20 l ice cream treatment group as well as 40 g/ml pure allergen compared with the PBS-treated group, and IL-4 cytokine levels decreased compared with the PBS-treated group. Overall, 20 l ice cream with 10 CFU/ml of the recombinant bacteria resulted in the best performance in terms of improving allergies to Th1 and Treg responses.
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http://dx.doi.org/10.1155/2020/2635230DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7532367PMC
September 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

Effect of experimental hyperthyroidism on and genes expression in the seminiferous tubules of BALB/c mice: An experimental study.

Int J Reprod Biomed 2020 Aug 19;18(8):591-604. Epub 2020 Aug 19.

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

Background: (Cation Channel Sperm Associated 1) and channels have an important role in sperm motility. In this study, the effects of hyperthyroidism on and genes of seminiferous tubules in mice testes were investigated.

Objective: The present study was conducted to investigate the effect of hyperthyroidism on the expression of and genes in the seminiferous tubules of mice.

Materials And Methods: This study was conducted on 20 BALB/C male mice divided into two groups - experimental and control. The experimental group was administered with 500 mg/l levothyroxine (L-thyroxine) liquid solution for two months for inducing hyperthyroidism, which was confirmed by radioimmunoassay. On the other hand, the control group was kept in animal houses under a normal condition. The implementation of real-time polymerase chain reaction and immunohistochemical studies was accomplished after the removal of the testes of the mice under anesthesia induced by chloroform.

Results: Results showed that there was no significant difference in (p = 0.45) and (p = 0.34) gene expression between groups. At the same time, the color intensity showed no significant enhancement in the hyperthyroidism group ( p = 0.17 and p = 0.22) as compared to the control group.

Conclusion: Considering the key role of in the molecular structure of the sperm, our findings showed that the hyperactivity of the thyroid gland has no significant effects on the function of these components. Therefore, it might be concluded that hyperthyroidism has no considerable effects on the seminiferous tubules.
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http://dx.doi.org/10.18502/ijrm.v13i8.7501DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7457153PMC
August 2020

Introducing a Stabilizer Formulation for Allergenic Mold Extracts.

Rep Biochem Mol Biol 2020 Apr;9(1):106-114

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

Background: Sensitization to common mold allergens is one of the major causes of allergic rhinitis and asthma. Therefore, there is a critical need for standard sensitivity tests including skin prick tests to improve the stability of fungi extracts in traditional allergenic formulations. To address this concern, the present study aimed to develop a formulation to preserve allergenic activity of mold extracts.

Methods: 48 stabilizer formulations were designed and monitored for allergenic activity during a 40-days incubation period at 37 °C using an ELISA. Specifically, the IgE reactivity of allergenic extracts were examined. After establishing the most effective stabilizer formulation, we evaluated whether it could protect the allergenic activity of Alt a1, , and using an IgE inhibition ELISA after 40 days at 37 °C.

Results: We demonstrated that the most effective stabilizer formulation was a glycerol-based extract containing Arg and Glu. This formulation had an equal ratio of sucrose, sorbitol and protein and was able to preserve more than 95% of allergenic extract activity during a 40-days incubation period at 37 °C.

Conclusion: The present study reveals a novel formulation that is an efficient stabilizer of allergenic mold extract activity and has practical applications in mold skin prick tests, ELISAs, immunotherapies, and RAST.
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http://dx.doi.org/10.29252/rbmb.9.1.106DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7424419PMC
April 2020

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

DC-targeted gold nanoparticles as an efficient and biocompatible carrier for modulating allergic responses in sublingual immunotherapy.

Int Immunopharmacol 2020 Sep 22;86:106690. Epub 2020 Jun 22.

Immuno-Biochemistry Lab, Immunology Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address:

Background: Sublingual immunotherapy (SLIT) was introduced to deliver allergens in an effective and non-invasive route, which can be considered as an alternative for allergen-specific subcutaneous immunotherapy (SCIT). On the other hand, the use of gold nanoparticles (AuNPs) in allergen delivery has beneficial effects on sublingual immunotherapy. In addition, the molecular targeting agents like aptamers (Apt), have been widely applied for targeted drug delivery. Therefore, the current study aimed to evaluate the effects of dendritic cells (DCs)-specific Aptamer-modified AuNPs coated with ovalbumin (OVA) on the improvement of the SLIT outcome in the mouse model of allergy.

Material And Methods: AuNPs with approximately 15 nm diameter were prepared by citrate reduction of HAuCl4. Afterward, Apt-modified AuNP complex was prepared and OVA was then loaded onto this complex. Following sensitization of Balb/c mice to OVA, SLIT was performed with Apt-AuNPs containing 5 µg OVA twice a week for a 2-month period. Allergen-specific IgE in serum, as well as cytokines secretion of spleen cells, were analyzed using ELISA. Also, nasopharyngeal lavage Fluid (NALF) was collected for total and eosinophil counts. Moreover, the lungs were removed for histopathological examination.

Results: SLIT with Apt-modified AuNPs complex containing 5 μg OVA, decreased the IgE levels compared to the other groups. Also, IL-4 production has significantly decreased in spleen cells, while TGF-β and IFN-γ have significantly increased. The assessment of NALF in the group treated by this complex showed a decrease in total cell as well as in eosinophil count. Also, the examination of lung tissues revealed that, in the group treated by this complex, inflammation and perivascular infiltration were lesser than the other groups, which were observed in only one vessel of tissue.

Conclusion: It was shown that, Sublingual immunotherapy with DC specific Apt-modified AuNPs containing 5 μg OVA can improve the Th1 and Treg immunomodulatory responses.
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http://dx.doi.org/10.1016/j.intimp.2020.106690DOI Listing
September 2020

Determining the Effect of Amino Acids on the Allergenic Activity of Pollen Extracts.

Rep Biochem Mol Biol 2020 Jan;8(4):394-400

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

Background: The diagnosis and treatment of allergic diseases require high quality pollen allergen extracts for reliable test results and effective treatments. The quality of the pollen allergen extracts is influenced by pharmacologically inert ingredients, such as stabilizers which are added to prevent the degradation of the allergenic activity. This study was conducted to develop a stabilizer formulation in order to protect the allergenic activity of the pollen's extracts.

Methods: Pine and orchard grass pollen allergen extracts were incubated for 40 days at 37 °C. The effects of chemicals were examined via inhibition ELISA on days 7, 14, 21, 28, and 40 to evaluate the ability of the pollen allergen extracts to inhibit specific IgE in the sera of sensitized patients.

Results: Our findings showed that the pine pollen and orchard grass allergen extracts treated with Lys/Glu had the best stabilizing effect resulting in a 97% IgE inhibition following the 40 days of incubation. In the non-treatment group, the IgE inhibition decreased to 23% at the end of the 40 days. The orchard grass pollen allergen extracts receiving no treatment decreased to 12% IgE inhibition following the 40-day incubation.

Conclusion: Amino acids are able to act as an effective stabilizer for pollen allergen extracts and prevent the degradation of their activity over time. Particularly applying Lys/ Glu in pollen allergenic extracts can protect allergenic activity and potency of the pollen extracts to inhibit specific IgE in human sera.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7275837PMC
January 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

Sublingual dendritic cells targeting by aptamer: Possible approach for improvement of sublingual immunotherapy efficacy.

Int Immunopharmacol 2020 Aug 30;85:106603. Epub 2020 May 30.

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

The efficacy improvement of current sublingual immunotherapy (SLIT) for preventing and treating respiratory airway allergic diseases is the main purpose of many investigations. In this study, we aimed to assess whether ovalbumin (Ova) encapsulated poly (lactic-co-glycolic) acid nanoparticles (PLGA NPs) decorated with dendritic cells (DCs)-specific aptamer could be applied for this purpose.The nanoparticles containing Ova were synthesized by emulsion/solvent evaporation method and attached to DCs-specific aptamer. Ova-sensitized BALB/c mice have been treated in five ways: subcutaneously with free Ova (SCIT), sublingually either with free Ova, Ova-PLGA NPs (two doses), Apt-Ova-PLGA NPs (two doses) and placebo/control Apt-Ova-PLGA NPs. For assessment of immunologic responses, IL-4, IFN-γ, IL-17, IL10, and TGF-β and IgE antibody levels were measured by ELISA and T cell proliferation were evaluated by MTT. In addition, lung and nasal histological examinations, NALF cells counting were carried out. Results declared that the lowest IgE and IL- 4 levels were observed in Apt-Ova-PLGA NPs (both doses). In the other hands, Apt-Ova-PLGA NPs (high dose) showed the highest increase of IFN- γ and TGF- β, decrease of IL-17 levels, total cell count and T-cell proliferation. IL-10 levels showed more decrease in SCIT, Apt-Ova-PLGA NPs (high dose) and Ova-PLGA NPs (high dose) than other groups. Histopathological examinations also confirmed in vitro results. Our findings suggest SLIT with this functionalized delivery system could be a promising approach for promoting the SLIT efficiency by decreasing the required allergen doses through specific delivery of allergen to sublingual DCs and enhancing the suppression of allergic responses.
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http://dx.doi.org/10.1016/j.intimp.2020.106603DOI Listing
August 2020

Dc-specific aptamer decorated gold nanoparticles: A new attractive insight into the nanocarriers for allergy epicutaneous immunotherapy.

Int J Pharm 2020 Jun 5;584:119403. Epub 2020 May 5.

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

Recently, the main goal of many allergy epicutaneous immunotherapy (EPIT) studies is to enhance the allergen delivery through the intact skin. Therefore, applying new strategies for tackling this issue are inevitable. For this purpose, ten groups of Che a 2-sensitized BALB/c mice were epicutaneously treated for a 6-week period with the rChe a 2-GNPs-Aptamer, rChe a 2-GNPs-Aptamer + skin-penetrating peptides (SPPs), rChe a 2-GNPs, rChe a 2, GNPs, and PBS. Afterward, the serum IgE and IFN-γ, TGF-β, IL-10, IL-4, IL-17a cytokine production, NALF analysis, and lung/nasal histological examinations were performed. The present study results demonstrate that, EPIT in aptamer treated groups had a significant increase of IFN-γ, TGF-β, and IL-10 concentrations and a significant decrease of IgE, IL-4, and IL-17a concentrations as well as NALF infiltrated immune cell count compared to the non-targeted ones. In addition, SPPs led to more significant improvement of immunoregulatory parameters, especially IL-10 cytokine. Accordingly, the targeted-GNPs with DC-specific aptamers could act as an efficient approach for the improvement of EPIT efficacy compared to the free allergen. Moreover, the application of SPPs might be considered as a useful tool in achieving a successful EPIT with lower doses of allergen at a shorter duration of the treatment.
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http://dx.doi.org/10.1016/j.ijpharm.2020.119403DOI Listing
June 2020

Production and Characterization of Monoclonal Antibody against Vit v1: A Grape Allergen Belonging to Lipid Transfer Protein Family.

Iran J Allergy Asthma Immunol 2020 Apr 16;19(2):139-148. Epub 2020 Apr 16.

Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Allergy to non-specific lipidtransfer protein (nsLTP), the major allergen of grape (Vit v1), is considered as one of the most common fruit allergies in Iran. Therefore, a specific monoclonal antibody (mAb) can be used for the characterization and assessment of. Accordingly, this study aimed to generate and characterize a mAb against Vit v1 with a diagnostic purpose. To this end, Vit v1 allergen (9 kDa) was extracted using a modified Bjorksten extraction method. Natural Vit v1-immunized mouse splenocytes were fused with SP2/0Ag-14 myeloma cells for generating hybridoma cells. Specific antibody-secreting Hybridoma cells were selected using ELISA. Finally, anti-Vit v1 mAb was characterized by western blotting, ELISA, and isotyping methods. In the current study, a 9 kDa (Vit v1) protein was attained fromcrude and fresh juice of grape extracts and the isotype of desired anti-Vit v1 mAb was determined as IgM with k light chain. In addition, The ELISA results demonstrated that anti-Vit v1 mAb was specified against natural Vit v1 in the grape cultivar and related LTP allergens, such as Pla or 3 (p<0.0001). In the present study, a specific mAb was produced for detecting the LTP allergen. This mAb with a confirmed specificity can be utilized for evaluating the LTP allergens and their allergenicity in different grape cultivars.
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http://dx.doi.org/10.18502/ijaai.v19i2.2776DOI Listing
April 2020

Immune responses modulation by curcumin and allergen encapsulated into PLGA nanoparticles in mice model of rhinitis allergic through sublingual immunotherapy.

Int Immunopharmacol 2020 Jul 28;84:106525. Epub 2020 Apr 28.

Nanotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad, University of Medical Sciences, Mashhad, Iran. Electronic address:

The purpose of this study was the combination of curcumin and ovalbumin in free form or encapsulated into PLGA NPs (polylactic co-glycolic acid nanoparticles) to enhance their sublingual immunotherapy (SLIT) efficiency in mouse model of rhinitis allergic. PLGA NPs containing curcumin (CUR), ovalbumin (OVA) or both were prepared by emulsion-solvent evaporation method and characterized. After sensitization of BALB/C mice with ovalbumin, SLIT with free or encapsulated formulations was carried out and immunological profiles were evaluated. SLIT treatment with all synthesized PLGA formulations lead to significantly decreased total IgE. The combination immunotherapy in the present of free form of curcumin or ovalbumin with encapsulated forms of the another substance (P.OVA-CUR 10 and P.CUR 5-OVA), showed the highest level of IFN-γ:IL-4 compared to other target groups. On the other hands, a significant increasment was observed in this ratio between these optimal groups and treated group with subcutaneous administration of OVA as the most commonly used method for immunotherapy. The study of nasal lavage fluid (NALF) showed significant decreased levels of total and eosinophil cell count in the traeted nano-formulation groups. The histopathological results of NAL were also like normal with no cellular infiltration and no inflammation in the optimal formulations. Therefore, using curcumin and nanoparticles with allergen can be considerd as potential immune modulatory agents.
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http://dx.doi.org/10.1016/j.intimp.2020.106525DOI Listing
July 2020

Designing a new dimerized anti human TNF-α aptamer with blocking activity.

Biotechnol Prog 2020 07 10;36(4):e2969. Epub 2020 Mar 10.

Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

The human tumor necrosis factor α (hTNF-α) is an important pro-inflammatory cytokine which plays critical roles in inflammatory diseases such as rheumatoid arthritis (RA). The anti-TNF-α proteins can reduce symptoms of RA. Due to limitations of protein-based therapies, it is necessary to find new anti-TNF-α agents instead of common anti-TNF-α proteins. Therefore, the aim of the current study was to identify a new DNA aptamer with anti-hTNF-α activity. The protein systematic evolution of ligands by exponential enrichment (SELEX) process was used for identifying DNA aptamers. Anti-hTNF-α aptamers were selected using dot blot, real-time PCR, and in vitro inhibitory assay. The selected aptamers were truncated in two steps, and finally, a dimer aptamer was constructed from different selected truncates to improve their inhibitory effect. Also, Etanercept was used as a positive control to inhibit TNF-α, in comparison to the designed aptamers. After 11 rounds, four aptamers with anti-hTNF-α inhibitory effect were identified. The truncation and dimerization strategy revealed a new dimer aptamer with 67 nM K , which has 40% inhibitory effect compared with Etanercept (60%). Overall, the dimerization and truncation aptamers could improve its activity. With regard to the several limitations of anti-TNF-α proteins therapies including immunogenicity, side effects, and cost-intensive, a new designed anti-hTNF-α dimer aptamer could be considered as a potential therapeutic and/or diagnostic agent for hTNF-α-related disorders.
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http://dx.doi.org/10.1002/btpr.2969DOI Listing
July 2020

Separation of the Epitopes in a Multi-Epitope Chimera: Helical or Flexible Linkers.

Protein Pept Lett 2020 ;27(7):604-613

Department of Immunology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Background: The engineered chimeric peptides including functional multi-epitope structures fused by various peptide linkers are widely applied in biotechnological research to improve the expression level and biological activity of chimera.

Objective: The aim of our study was to evaluate the effect of helical and flexible linkers on solubility, expression level and folding of multi-epitope chimera containing four epitopes of Human T Lymphotropic Virus Type 1 (HTLV-1).

Methods: For this purpose, the chimera sequences connected by the helical or flexible linker were inserted into different plasmid vectors and expressed in E. coli strains. The expressed products were analyzed using SDS-PAGE and Western blot techniques. Additionally, the molecular modeling study of the chimera with helical or flexible linker was performed using iterative threading assembly refinement (I-TASSER) to attain their three-dimensional structures.

Results: Comparison of the chimera expression indicated that the insertion of a flexible (GGGGS)3 linker among chimera epitopes could significantly enhance the level of expression, whereas, the low-level of chimera expression was observed for chimera containing the contiguous helical (EAAAK)5 linker. According to the results of sequence alignment and plasmid stability test, the structure and function of a consecutive helical linker among chimera epitopes were similar to porins as the outer-membrane pore-forming proteins. The molecular modeling results confirmed our experimental study.

Conclusion: This investigation illustrated the key role of linker design in determining the expression level of multi-epitope chimera and conformational folding.
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http://dx.doi.org/10.2174/0929866526666191112124602DOI Listing
January 2021

Analysis of different signal peptides for the secretory production of Ama r 2 in gram-positive systems (Lactococcus lactis).

Microb Pathog 2020 Jan 25;138:103819. Epub 2019 Oct 25.

Department of Food Science and Technology, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.

Prokaryotic systems have been considered the most affordable and simplest hosts which are being employed to express recombinant proteins such as allergens; nevertheless, without appropriate signal peptide (SP), these systems cannot be used for secretory proteins. Recently, a lot of effort has been put into assessing the potential of gram-positive strains such as lactic acid bacteria for new applications in the production of heterologous proteins. Ama r 2 is a respiratory allergen from Amaranthus retroflexus, whose recombinant production in the probiotic host could be introduced as a specific and effective way to rapid diagnosis and immunotherapy of this allergy. Consequently, the production of this recombinant protein using the prokaryotic system, requires a suitable SP to protect disulfide bonds and to prevent misfolding. This study was designed to predict the best SPs for the expression of Ama r 2 protein in Lactococcus lactis as the host. In this study, 42 signal sequences were selected from SP databases and the most important features of them were evaluated. First, n, h and c regions of the SPs and their probabilities were investigated by signalP software version 4.1. Then, their physicochemical properties were evaluated by Portparam and SOLpro. Moreover, the secretion sorting and sub-cellular localization sites were evaluated by PRED-TAT and ProtcompB software programs. The results revealed that yjgB, entC2 (Entrotoxine type C-2), ent B (Entrotoxine type), blaZ (Beta lactamase), dex (number 21), blm (Beta lactamase 2), dex (Dextranase; number 20) and number 26 were introduced theatrically as the best SPs to express Ama r 2 in Lactococcus lactis.
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http://dx.doi.org/10.1016/j.micpath.2019.103819DOI Listing
January 2020

The immunotoxin activity of exotoxin A is sensitive to domain modifications.

Int J Biol Macromol 2019 Aug 23;134:1120-1131. Epub 2019 May 23.

Department of Medicinal Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran; Bioinformatics Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address:

Immunotoxins are a class of recombinant proteins which consist of an antibody and a part of a bacterial or herbal toxin. Immunotoxins containing Pseudomonas aeruginosa exotoxin A (PEA) have been found to be very applicable in clinical trials. Many obstacles such as solubility and absorbency reduce their usability in solid tumors. The current study aims to overcome the mentioned barriers by addition and removal of functional and non-functional domains with a structural approach. In the experimental section, we took advantage of molecular dynamics simulations to predict the functionality of candidate immunotoxins which target human HER2 receptors and confirmed our findings with in vitro experiments. We found out when no changes were made to domain II of PEA, addition of solubilizing domains to immunotoxins would not reduce their targeting and anti-tumor activity, while increasing the yield of expression and stability. On the other side, when we replaced domain II with eleven amino acids of furin cleavage site (FCS), the activity of the immunotoxin was mainly affected by the FCS neighboring domains and linkers. A combination of seven beneficial point mutations in domain III was also assessed and reconfirmed that the toxicity of the immunotoxin would be reduced dramatically. The obtained results indicate that the addition or removal of domains cannot depict the activity of immunotoxins and the matter should be assessed structurally in advance.
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http://dx.doi.org/10.1016/j.ijbiomac.2019.05.137DOI Listing
August 2019

Antagonistic activity of recombinant Lactococcus lactis NZ1330 on the adhesion properties of Escherichia coli causing urinary tract infection.

Microb Pathog 2019 Aug 18;133:103547. Epub 2019 May 18.

Department of Food Science and Technology, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.

Death from infectious diseases has caused concerns about increases in the resistance of pathogens, impelling researchers to create novel therapeutic solutions. The management of intestinal tract problems has been the advance use of probiotics in medicine. The aim of this study was evaluate the physicochemical cell surface and adhesion properties of recombinant Lacotococcus lactis NZ1330 containing Ama r 2 gene, followed by the assessment of the antagonistic activity of this strain against the Escherichia coli causing urinary tract infection (UTI) in humans. For this purpose, cloning and expression of Ama r 2 gene were done. Afterwards, acid and bile resistance, which are the primary characteristics of any probiotic, were evaluated. The r-L. lactis NZ1330 was examined for the physicochemical properties of cell surfaces and the adhesion properties against Escherichia coli. Furthermore, the potential of the recombinant strain to adhere to adenocarcinoma intestinal cell line, Caco-2 cells, as well as the antagonistic properties of r-L. lactis NZ1330 against E. coli was investigated. r-L. lactis NZ1330 was capable of surviving at low pH and different concentrations of bile salts. 40.1% hydrophobicity, 36.5% auto-aggregation and 14.4% co-aggregation were observed for this strain. The adhesion level of r-L. lactis NZ1330 was 5.7% which was also confirmed by scanning electron microscopy (SEM). r-L. lactis NZ1330 was able to compete, inhibit and displace the adhesion of Escherichia coli to Caco-2 cells. r-L. lactis NZ1330 was considered to be a reliable probiotic alternative by showing these desirable properties. Results revealed that Ama r 2 gene expression had no effect on the positive probiotic properties of L. lactis NZ1330, proving this strain could be a suitable probiotic host for the expression of this allergen.
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http://dx.doi.org/10.1016/j.micpath.2019.103547DOI Listing
August 2019

The Most Common Allergenic Tree Pollen Grains in the Middle East: A Narrative Review.

Iran J Med Sci 2019 Mar;44(2):87-98

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

Allergy is becoming a major disease burden globally. Pollens are considered as the main component of aeroallergens that lead to rhinitis and asthma. Due to the lack of a comprehensive investigation on most allergic pollens of trees in the Middle East, the present study aimed to conduct a comprehensive literature review on this topic. The main goal of the study was to provide a checklist for allergists and patients to easily identify the commonest allergic pollens in their locality. The present review provides a broad range of information on the types and geographic locations of the most common allergic pollens of trees in each studied country. In general, among the 23 studied countries, palm and mesquite trees were the common producers of pollen allergen in the Persian Gulf region. Olive tree is common in Turkey, Palestine, and Israel, whereas sycamore tree is the common allergen pollen in Iran. Considering the uneven geographical distribution of these trees in the world, allergists are unable to accurately select the appropriate extracts for the skin prick test based on the information from the neighboring countries. This scenario becomes more complicated if one adds the imported ornamental trees in the picture.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6423439PMC
March 2019

Evaluation of size and dose effects of rChe a 3 allergen loaded PLGA nanoparticles on modulation of Th2 immune responses by sublingual immunotherapy in mouse model of rhinitis allergic.

Int J Pharm 2019 May 19;563:282-292. Epub 2019 Mar 19.

Immunology Research Center, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address:

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http://dx.doi.org/10.1016/j.ijpharm.2019.03.040DOI Listing
May 2019

Determination of Optimum Excipients for Platanus orientalis Pollen Extract by Accelerating Chemical Stability Test and Their Synergistic Effect.

Rep Biochem Mol Biol 2019 Jan;7(2):189-195

Allergy Research Center, Ghaem Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Background: The quality of extracts used in the skin prick test directly influences the interpretation of the test. Accordingly, the outcomes and effectiveness of immunotherapy for the management of IgE-mediated allergies depend on the quality of the extracts used. Excipients, which are pharmacologically inert ingredients, are intentionally added to the active ingredients. The aim of this study was to address optimum excipients for stability extract.

Methods: In this study the excipients examined were l-lysine (20 mM), l-cysteine (20 mM), albumin (0.5%), sorbitol (2%), sucrose (750 mM), trehalose (20 mM), D-mannitol (2% w/v), urea (100 mM) and Tween-20 (0.1%). Their effects on P. orientalis extract stability were analyzed using an inhibition enzyme linked immune assay at 37 °C.

Results: A mixture of lysine (20 mM), trehalose (20 mM), and D-mannitol (2% w/v) conferred the greatest stability on the extract.

Conclusion: The extract stability was increased by a mixture of lysine (20 mM), trehalose (20 mM), and D-mannitol.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6374054PMC
January 2019

The Influence of Gamma Radiation Processing on the Allergenicity of Main Pistachio Allergens.

Rep Biochem Mol Biol 2019 Jan;7(2):150-155

Immunology Research Center, Bu-Ali Research Institute, University of Medical Sciences, Mashhad, Iran.

Background: Gamma irradiation is a form of processing with an array of applications in medical sciences such as microbial decontamination, viruses inactivation, cervical carcinoma and breast cancer treatment. One of the ways in which gamma irradiation has the potential to be used is in reducing the allergenicity of food allergens.

Methods: In the present study, pistachios were irradiated with either a 1, 10, or 100 kGy dose of gamma irradiation. The binding rate of mice and human antibodies to the allergens of the pistachio extracts were examined via Western blot analysis.

Results: Our findings show an inverse dose-response relationship between the binding rate of antibodies to the pistachio allergens and the gamma irradiation dose. Despite these promising findings, the results of our sensory evaluation indicate that gamma irradiation causes undesirable changes to the sensory characteristics of pistachios, especially at the dose of 100 kGy.

Conclusion: Gamma irradiation appears to be an effective method in reducing the allergenicity of pistachios. Thus, this form of processing has the potential to prevent adverse allergic reactions to the major pistachio allergens in sensitized subjects. However, further research must be dedicated to examining the dose sufficient in reducing allergencity, while maintaining adequate sensory quality for satisfactory consumption.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6374055PMC
January 2019

Mutation analysis of genes related to methylmalonic acidemia: identification of eight novel mutations.

Mol Biol Rep 2019 Feb 2;46(1):271-285. Epub 2019 Feb 2.

Department of Medical Laboratory Sciences, Varastegan Institute for Medical Sciences, Mashhad, Iran.

Methylmalonic acidemia (MMA), an inherited metabolic disease, results from genetic defects in methylmalonyl-CoA mutase or any of the proteins involved in adenosylcobalamin synthesis. This enzyme is classified into several complementation groups and genotypic classes. In this work we explain the biochemical, structural and genetic analysis of 25 MMA patients, from Iran. The diagnosis was established by the measurement of propionylcarnitine in blood using tandem mass spectrometry and confirmed using a gas chromatography-flame ionization detector. Using clinical, biochemical, structural and molecular analyses we identified 15 mut MMA, three cblA, one cblB, and four cblC-deficient patients. Among mutations identified in the MUT gene (MUT) only one, the c.1874A>C (p.D625A) variant, is likely a mut mutation. The remaining mutations are probably mut. Here, we present the first molecular analysis of MMA in Iranian patients and have identified eight novel mutations. Four novel mutations (p.D625A, p.R326G, p.V157F, p.F379L) were seen exclusively in patients from northern Iran. One novel splice site mutation (c.2125-3C>G) in MUT and two novel mutation (p.N225M and p.A99P) in the MMAA gene were associated with patients from eastern Iran. The rs184829210 SNP was recognized only in patients with the novel c.958G>A (p.A320T) mutation. This study confirms pathogenesis of deficient enzyme activity in MUT, MMAA, MMAB, and MMACHC as previous observations. These results could act as a basis for the performance of pharmacological therapies for increasing the activity of proteins derived from these mutations.
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http://dx.doi.org/10.1007/s11033-018-4469-0DOI Listing
February 2019