Publications by authors named "Fatemeh Khademi"

27 Publications

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Selenium nanoparticles: Synthesis, cytotoxicity, antioxidant activity and interaction studies with ct-DNA and HSA, HHb and Cyt c serum proteins.

Biotechnol Rep (Amst) 2021 Jun 15;30:e00615. Epub 2021 Apr 15.

Center of Medical Biology Research, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.

The aim of this study was the synthesis of selenium nanoparticles (SeNPs) employing vitamin C as a biocompatible and low toxic reducing agent. The synthesized selenium nanoparticles were characterized by using UV-vis, FT-IR, SEM-EDX, TEM, DLS, and zeta potential measurements. The results of the DPPH free radical scavenging assay demonstrate that this synthesized nano-selenium has strong potentials to scavenge the free radicals and cytotoxicity against MCF-7 and Raji Burkitt's lymphoma cancer cell lines. The interaction of calf thymus DNA (ct-DNA) with SeNPs indicated that the anticancer activity might be associated with the DNA-binding properties of nano-selenium. Finally, it was found that the synthesized nano-selenium can bind to the most important blood proteins such as human serum albumin (HSA), human hemoglobin (HHb), and Cytochrome c (Cyt c). The results showed that the secondary structure of these proteins remains unchanged, suggesting that the synthesized nano-selenium could be employed as a carrier in the drug delivery system without any cytotoxicity effect.
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http://dx.doi.org/10.1016/j.btre.2021.e00615DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8080047PMC
June 2021

The Construction and Characterization of 1F5 Chimeric Anti-CD20 Monoclonal Antibodies: An Efficient Splicing by Overlap Extension-polymerase Chain Reaction Technique.

Iran J Allergy Asthma Immunol 2021 Apr 17;20(2):205-220. Epub 2021 Apr 17.

Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran.

Despite the unparalleled success of anti-CD20-targeted immunotherapy, the currently available mAbs are not sufficiently efficacious in the treatment of lymphoma. 1F5 is one of a panel of anti-CD20 mAbs that was used in the B-cell lymphoma serotherapy. Despite the efficacy of murine 1F5 mAbs in lymphoma patients, the 1F5 chimeric antibodies with human effector functionality are yet to be approved and widely used in the treatment of lymphoma. In this study, the conversion of 1F5 mAb from mouse IgG2a to human-mouse chimeric IgG1 was achieved and the chimeric antibody was partially characterized. We constructed the 1F5 chimeric mouse-human anti-CD20 antibody genes using an efficient Splicing by overlap extension-polymerase chain reaction (SOE-PCR) technique and cloned the chimeric heavy and light genes in pBudCE4.1mammalian expression vector, followed by purification of the expressed chimeric 1F5 mAbs using affinity chromatography. Our investigation also included the biological properties of purified chimeric antibodies. The generated 1F5 chimeric mAbs mediate complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC) against Raji and Daudi Burkitt's lymphoma cell lines, which were comparable with rituximab and exhibit superior reduction in cell viability in vitro, compared to rituximab. The current study indicated that the generated chimeric 1F5 mAbs has potential CDC and ADCC activity which was comparable with rituximab whereas it exhibits a superior reduction in cell viability, compared to rituximab. Our work contributes to future studies involving in vivo biological functions and the application of the 1F5 chimeric antibody.
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April 2021

Psychometric Properties of the Farsi Version of Posttraumatic Growth Inventory for Children-Revised in Iranian Children with Cancer.

Asia Pac J Oncol Nurs 2021 May-Jun;8(3):295-303. Epub 2021 Mar 26.

Department of Pediatric and Neonatal Intensive Care Nursing, School of Nursing and Midwifery, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Objective: Coping with childhood cancer, as a stressful incident, can lead to a growth in various aspects of the child's life. Therefore, this study aims to validate Posttraumatic Growth Inventory for Children-Revised (PTGI-C-R) in children with cancer.

Methods: This methodological research was carried out in referral children hospitals in Tehran. PTGI-C-R was translated and back-translated. Content and face validity were assessed. Confirmatory factor analysis (CFA) was performed on 200 children with inclusion criteria, using LISREL V8.5. Due to the rejection of the model, an exploratory factor analysis (EFA) was done, using SPSS V21. The correlation of posttraumatic growth (PTG) with the variables, i.e., age and gender, was investigated.

Results: Some writing changes were made in phrases in the sections concerning face and content validity. CFA rejected the five-factor model due to the undesirable fit indices. Therefore, an EFA was used and the three-factor model was not approved, either despite the statistical appropriateness or due to the lack of similarity between the items loaded on factors. The results also indicated a significant relationship between PTG and age ( = 0.13, = 0.05). There is no significant relationship between PTG and gender ( = -1.35, = 0.83).

Conclusions: PTGI-C-R does not have desirable psychometric properties in Iranian children with cancer and may not be able to reflect all the aspects of PTG experienced by them. Therefore, it cannot be used as an appropriate scale, and it is necessary to develop and validate a specific tool through a qualitative study.
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http://dx.doi.org/10.4103/apjon.apjon-2093DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8030598PMC
March 2021

The Antimicrobial Peptide, Nisin, Synergistically Enhances the Cytotoxic and Apoptotic Effects of Rituximab Treatment on Human Burkitt's Lymphoma Cell Lines.

Rep Biochem Mol Biol 2020 Oct;9(3):250-256

Medical Biology Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.

Background: Non-Hodgkin's lymphomas comprise the most common hematological cancers worldwide and consist of a heterogenous group of malignancies affecting the lymphoid system. Treatment of non-Hodgkin's lymphoma has been significantly enhanced with the addition of Rituximab to the standard chemotherapy regimen. However, even with the advancement of treatment patients continue to relapse and develop resistance to Rituximab, rendering subsequent treatments unsuccessful. The use of novel drugs with unique antitumor mechanisms has gained considerable attention. In this study, we explored the anti-cancer effects of the combined therapy of Rituximab and Nisin on human Burkitt's lymphoma cells.

Methods: The human Burkitt's lymphoma cells lines, Raji and Daudi, were treated with Nisin, Rituximab, or a combination of the two agents at various concentrations. Cytotoxicity following treatment was determined using cell viability assay. The degree of apoptosis was verified via flow cytometric analysis using FITC annexin V/PI staining.

Results: Our findings show that the combined treatment of Rituximab and Nisin results in a more significant reduction in the survival of Raji and Daudi Burkitt's lymphoma cells, compared to Nisin or Rituximab treatment alone. Additionally, our results indicate that Nisin can induce a significant degree of apoptosis in the Burkitt's lymphoma cells compared to the negative controls. However, the addition of Nisin to the Rituximab treatment synergistically enhances the apoptotic antitumor effect.

Conclusion: This study demonstrates the synergistic antitumor effect of Nisin treatment to enhance tumor cell apoptosis and the potential value of Nisin as an adjunct therapy in the treatment of lymphoma.
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http://dx.doi.org/10.29252/rbmb.9.3.250DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7816786PMC
October 2020

The COVID-19 pandemic and death anxiety in the elderly.

Int J Ment Health Nurs 2020 Dec 1. Epub 2020 Dec 1.

Department of Nursing, School of Nursing, Arak University of Medical Sciences, Arak, Iran.

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http://dx.doi.org/10.1111/inm.12824DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7753683PMC
December 2020

Psychometric Evaluation of the Caring Ability of Family Caregivers of Patients With Cancer Scale-Mothers' Version for the Mothers of Children With Cancer.

Cancer Nurs 2020 Oct 30. Epub 2020 Oct 30.

Author Affiliations: Department of Nursing, Faculty of Nursing, Arak University of Medical Sciences, Arak (Ms Khademi); Department of Pediatric Nursing, School of Nursing and Midwifery (Dr Rassouli); Department of Medical Surgical Nursing, School of Nursing and Midwifery (Dr Khanali Mojen); and Department of Pediatric Nursing, School of Nursing and Midwifery (Dr Shirinabadi Farahani), Shahid Beheshti University of Medical Sciences, Tehran; and Department of Nursing, School of Nursing and Midwifery, Ardabil University of Medical Sciences, Ardabil, Iran (Dr Heidarzadeh).

Background: The experience of caring for cancer patients has adverse outcomes for family caregivers. The ability to care for a sick child is affected by the mother's health; to empower mothers, it will be necessary to examine their caring ability.

Objective: The aim of this study was to carry out a psychometric evaluation of the Caring Ability of Family Caregivers of Patients With Cancer Scale-Mothers' Version (CAFCPCS-Mothers' Version).

Methods: The present study is a psychometric evaluation of the CAFCPCS-Mothers' Version. The sample consisted of 196 mothers of children in treatment for cancer selected through convenience sampling. The face, content and construct validity, internal consistency, and stability of the scale were measured. Data were analyzed using the software SPSS 19 and LISREL 8.8.

Results: After removing 2 items during confirmatory factor analysis, the values of root-mean-square error of approximation, comparative fit index, and nonnormed fit index were reported to be 0.066, 0.92, and 0.91, respectively. The Cronbach's α was calculated to be 0.71 and the stability correlation coefficient was 0.75. The final tested scale included 29 items in 5 dimensions: effective role play, fatigue and surrender, trust, uncertainty, and caring ignorance for mothers of children with cancer.

Conclusion: The CAFCPCS-Mothers' Version has satisfactory content, face, and construct validity and adequate reliability in terms of internal consistency and stability in a sample of mothers of children receiving treatment for cancer.

Implication For Practice: The CAFCPCS-Mothers' Version can be used to assess the caring ability of Iranian mothers of children with cancer and to determine maternal care needs.
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http://dx.doi.org/10.1097/NCC.0000000000000902DOI Listing
October 2020

Production of monoclonal antibody against recombinant bovine sex-determining region Y (SRY) and their preferential binding to Y chromosome-bearing sperm.

Reprod Domest Anim 2021 Feb 3;56(2):270-277. Epub 2021 Jan 3.

Medical Biology Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.

Separation of X and Y chromosome-bearing sperm is an appropriate method for the selection of desired sex of offspring to increase the profit in livestock industries. The purpose of this study was the production of a monoclonal antibody against recombinant bovine sex-determining region Y protein for separation Y sperm. The hybridoma cells from splenocytes of immunized female's balb/C mice and Sp2/0 cells were made. The binding affinity of our monoclonal antibody (mAbSRY2) was compared with mouse monoclonal SRY-15. The Western blot method indicated that mAbSRY2 successfully detected the rbSRY protein. The specificity and sensitivity of mAbSRY2 is comparable to SRY-15 commercially ones. The SRY gene in 100% of bull semen contains the Y chromosome that had the strongest binding affinity to mAbSRY2 was synthesized. In other words, the binding affinity of semen contains the X sperms near the negative control. In general, this immunological method can help to separate X from Y sperms. However, the mAbSRY2 is bind to Y-bearing sexed sperm, but in the future; the sexed sperms need to apply in farms.
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http://dx.doi.org/10.1111/rda.13821DOI Listing
February 2021

Glutathione-dependent enzymes in the follicular fluid of the first-retrieved oocyte and their impact on oocyte and embryos in polycystic ovary syndrome: A cross-sectional study.

Int J Reprod Biomed 2020 Jun 30;18(6):415-424. Epub 2020 Jun 30.

Infertility Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.

Background: Oxidative stress and GSH-dependent antioxidant system plays a key role in the pathogenesis of polycystic ovary syndrome (PCOS).

Objective: We compared glutathione peroxidase (GPx) and glutathione reductase activities and reduced glutathione (GSH) levels in serum and follicular fluid (FF) of the first-retrieved follicle and their impact on quality of oocyte and embryo in PCOS women undergoing IVF.

Materials And Methods: This cross sectional study was conducted on 80 pairs of blood samples and FF of the first-retrieved follicle from PCOS women, at the Infertility center of Ghadir Mother and Child Hospital. The mean activity of GPx and GR, also GSH levels in the serum and FF were compared to the quality of the first follicle and resultant embryo.

Results: Retrieved oocytes included 53 (66.25%) MII, 17 (21.25%) MI, and 10 (12.5%) germinal vesicles; after IVF 42 (52.50%) embryos with grade I and 11 (13.75%) with grade II were produced. The mean values for all three antioxidants were higher in the FF compared to serum (p 0.001). Also all of the mean measured levels were significantly higher in the FF of the MII oocytes compared to that of oocytes with lower grades (p = 0.012, 0.006 and 0.012, respectively). The mean GPX activity and GSH levels were significantly higher in the serum (p = 0.016 and 0.012, respectively) and FF (p = 0.001 for both) of the high-quality grade I embryos.

Conclusion: GSH-dependent antioxidant system functions more efficiently in the FF of oocytes and embryos with higher quality.
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http://dx.doi.org/10.18502/ijrm.v13i6.7283DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7340988PMC
June 2020

Visual feature tuning of superior colliculus neural reafferent responses after fixational microsaccades.

J Neurophysiol 2020 06 29;123(6):2136-2153. Epub 2020 Apr 29.

Werner Reichardt Centre for Integrative Neuroscience, Tuebingen University, Tuebingen, Germany.

The primate superior colliculus (SC) is causally involved in microsaccade generation. Moreover, visually responsive SC neurons across this structure's topographic map, even at peripheral eccentricities much larger than the tiny microsaccade amplitudes, exhibit significant modulations of evoked response sensitivity when stimuli appear perimicrosaccadically. However, during natural viewing, visual stimuli are normally stably present in the environment and are only shifted on the retina by eye movements. Here we investigated this scenario for the case of microsaccades, asking whether and how SC neurons respond to microsaccade-induced image jitter. We recorded neural activity from two male rhesus macaque monkeys. Within the response field (RF) of a neuron, there was a stable stimulus consisting of a grating of one of three possible spatial frequencies. The grating was stable on the display, but microsaccades periodically jittered the retinotopic RF location over it. We observed clear short-latency visual reafferent responses after microsaccades. These responses were weaker, but earlier (relative to new fixation onset after microsaccade end), than responses to sudden stimulus onsets without microsaccades. The reafferent responses clearly depended on microsaccade amplitude as well as microsaccade direction relative to grating orientation. Our results indicate that one way for microsaccades to influence vision is through modulating how the spatio-temporal landscape of SC visual neural activity represents stable stimuli in the environment. Such representation depends on the specific pattern of temporal luminance modulations expected from the relative relationship between eye movement vector (size and direction) on one hand and spatial visual pattern layout on the other. Despite being diminutive, microsaccades still jitter retinal images. We investigated how such jitter affects superior colliculus (SC) activity. We found that SC neurons exhibit short-latency visual reafferent bursts after microsaccades. These bursts reflect not only the spatial luminance profiles of visual patterns but also how such profiles are shifted by eye movement size and direction. These results indicate that the SC continuously represents visual patterns, even as they are jittered by the smallest possible saccades.
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http://dx.doi.org/10.1152/jn.00077.2020DOI Listing
June 2020

Nicotine-Induced Oxidative Stress in Human Primary Endometrial Cells.

Int J Toxicol 2019 May/Jun;38(3):202-208. Epub 2019 May 21.

1 Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Nicotine is a major component of tobacco plants and is responsible for the development of reproductive problems in smokers. Nicotine has been recognized to result in oxidative stress by inducing the generation of reactive oxygen species (ROS) in some parts of female reproductive system, but the effect of nicotine on endometrium that plays an important role in reproductive biology stays unexplored. The aim of this work was to clarify the direct effects of nicotine administration on the antioxidant defense system and lipid peroxidation in human endometrial cells. Human endometrial stromal primary cells were treated with nicotine (0, 10, 10, and 10 M) for 24 hours. On nicotine administration, the endometrial cells were associated with a decrease in antioxidant defense markers such as Glutathione (GSH) level, glutathione peroxidase (GPx), glutathione reductase (GR), and catalase (CAT) enzymes activity and higher levels of malondialdehyde (MDA) in a dose-dependent manner when compared to the control. We concluded that nicotine as a pro-oxidant affects the oxidative state of the endometrial cells.
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http://dx.doi.org/10.1177/1091581819848081DOI Listing
January 2020

State-dependent brain stimulation: Power or phase?

Brain Stimul 2019 Mar - Apr;12(2):296-299. Epub 2018 Oct 26.

Division of Functional and Restorative Neurosurgery, and Tuebingen NeuroCampus, Eberhard Karls University Tuebingen, Germany. Electronic address:

Background: Intrinsic motor cortex activity modifies corticospinal excitability (CSE) in accordance with both oscillatory power fluctuations and phase-specific modulation along the oscillatory beta cycle, particularly in the 16-17 Hz frequency bin.

Objective: To determine the magnitude of CSE and the relevance of stimulation timing for input gain mediated by either oscillatory power or phase.

Methods: We applied single-pulse transcranial magnetic stimulation (TMS) over the primary motor cortex of healthy subjects at rest during electroencephalography recordings. The corticospinal gain modulation was indexed by the amplitude variability of the induced motor-evoked potentials recorded from the forearm muscle.

Results: Low compared to high beta power led to a robust 40-70% CSE increase over a wide range of power values. By contrast, the phase modulation was critically dependent on the precise timing of the stimuli to the rising phase of the oscillatory beta cycle, but could then achieve CSE increases of 180%.

Conclusion: These findings can influence closed-loop, state-dependent stimulation in the context of neurorehabilitation.
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http://dx.doi.org/10.1016/j.brs.2018.10.015DOI Listing
June 2019

In-Vitro Analysis of Glucose and Quercetin Effects on m-TOR and Nrf-2 Expression in HepG2 Cell Line (Diabetes and Cancer Connection).

Nutr Cancer 2018 07 21;70(5):770-775. Epub 2018 May 21.

a Biochemistry Department , School of Medicine, Shiraz University of Medical Sciences , Shiraz , Iran.

Some types of cancers show a strong relationship with diabetes and play a central role in mortality in the patient population suffering from diabetes mellitus. In this study, HepG2 cells have been used to investigate the toxic effects of hyperglycemia and/or quercetin (Q) on mammalian target of rapamycin (m-TOR) and nuclear factor erythroid 2-related factor 2 (Nrf-2) expression as central molecules involved in cancer. HepG2 cells were cultured with different concentrations of glucose (5.5, 30, and 50 mM) and/or Q (25 µM) for 48 and 72 h. Effects of glucose and/or Q on m-TOR and Nrf-2 expression were assayed by quantitative real-time PCR (qRT-PCR). qRT-PCR results revealed that 30 and 50 mM of glucose increased m-TOR expression at 48 h, although after 72 h, only 30 mM had an increasing effect. At 50 mM, glucose-induced Nrf-2 gene expression after both 48 and 72 h. The results also showed that 25 µM of Q reduced m-TOR and Nrf-2 expression at both 30 and 50 mM after 48 and 72 h incubation. Q has potential effects on reducing oxidative stress caused by hyperglycemia and during diabetes may be able to modulate some carcinogenic signaling pathways.
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http://dx.doi.org/10.1080/01635581.2018.1470654DOI Listing
July 2018

Distinct Beta-band Oscillatory Circuits Underlie Corticospinal Gain Modulation.

Cereb Cortex 2018 04;28(4):1502-1515

Division of Functional and Restorative Neurosurgery, and Centre for Integrative Neuroscience, Eberhard Karls University Tuebingen, 72076 Tuebingen, Germany.

Rhythmic synchronization of neurons is known to affect neuronal interactions. In the motor system, oscillatory power fluctuations modulate corticospinal excitability. However, previous research addressing phase-specific gain modulation in the motor system has resulted in contradictory findings. It remains unclear how many time windows of increased responsiveness each oscillatory cycle provides. Moreover, we still lack conclusive evidence as to whether the motor cortex entails an intrinsic response modulation along the rhythm cycle, as shown for spinal neurons. We investigated this question with single-pulse transcranial magnetic stimulation over the primary motor cortex at rest. Application of near-motor threshold stimuli revealed a frequency- and phase-specific gain modulation at both cortical and spinal level, independent of the spontaneous oscillatory power fluctuations at each level. We detected bilateral sensorimotor circuits in the lower beta-band (14-17 Hz) and unilateral corticospinal circuits in the upper beta-band (20-24 Hz). These findings provide novel evidence that intrinsic activity in the human motor cortex modulates input gain along the beta oscillatory cycle within distinct circuits. In accordance with periodic alternations of synchronous hyper- and depolarization, increased neuronal responsiveness occurred once per oscillatory beta cycle. This information may lead to new brain state-dependent and circuit-specific interventions for targeted neuromodulation.
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http://dx.doi.org/10.1093/cercor/bhy016DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6093341PMC
April 2018

An Evidence-Based Study on Medicinal Plants for Hemorrhoids in Medieval Persia.

J Evid Based Complementary Altern Med 2017 10 24;22(4):969-981. Epub 2017 Jan 24.

4 Medicinal Plants Processing Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.

Hemorrhoids is one of the most common gastrointestinal diseases. There are several therapeutic options associated with some complications. Therefore, researchers look for traditional medicines as a potential resource for introduction of new natural drugs. The current study reports an evidence-based review of herbal remedies for hemorrhoids in traditional Persian medicine. A comprehensive survey about hemorrhoids on the most important manuscripts of traditional Persian medicine was done. Then, scientific data banks were searched for possible related properties of each herb in the conventional medicine. We reported some historical aspects of traditional Persian medicine view on classification, examination, and predisposing factors of hemorrhoids. In addition, we have reported 105 medicinal plants belonging to 51 families. More than half of the reported herbs exhibited anti-inflammatory and analgesic effects. Although lack of human studies regarding the mentioned herbs is noted, positive results from experimental findings can be considered for new drug discovery supported by traditional and medieval experiences.
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http://dx.doi.org/10.1177/2156587216688597DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5871264PMC
October 2017

Oral Contraceptive Use May Modulate Global Genomic DNA Methylation and Promoter Methylation of APC1 and ESR1

Asian Pac J Cancer Prev 2017 09 27;18(9):2361-2366. Epub 2017 Sep 27.

Biochemistry and Genetics Department, Faculty of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran. Email:

Background: There are challenging reports in the public health sphere regarding associations between oral contraceptive (OC) use and cancer risk. Methods: To evaluate possible effects of OCs on cancer susceptibility, we quantified of global 5-methyl cytosine (5-mC) levels and assessed methylation patterns of CpG islands of two key tumor suppressor genes, APC1 and ESR1, in serum of users by enzyme-linked immunosorbent assay and methylation specific PCR methods, respectively. Results: Our results indicated that OCs significantly decrease the level of global DNA methylation in users relative to control non-users. However, our data revealed no significant differences between CpG island methylation patterns for ESR1 and APC1 in healthy control and OC-treated women. However, we did find a trend for hypermethylation of both tumor suppressor genes in OC users. Conclusion: Our data suggest that the level of 5-mC but not individual CpG island patterns is significantly influenced by OCs in our cross-section of adult users.
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http://dx.doi.org/10.22034/APJCP.2017.18.9.2361DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5720637PMC
September 2017

Construction and characterization of monoclonal antibodies against the extracellular domain of B-lymphocyte antigen CD20 using DNA immunization method.

Int Immunopharmacol 2017 Feb 7;43:23-32. Epub 2016 Dec 7.

Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran; School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran.

To date, several new anti-CD20 monoclonal antibodies (mAbs) have been developed for potential efficacies compared with familiar mAb rituximab. Despite the recent advances in development of anti-CD20 mAbs for the treatment of B cell malignancies, the efforts should be continued to develop novel antibodies with improved properties. However, the development of mAbs against CD20 as a multi-transmembrane protein is challenging due to the difficulty of providing a lipid environment that can maintain native epitopes. To overcome this limitation, we describe a simple and efficient DNA immunization strategy for the construction of a novel anti-CD20 mAb with improved anti-tumour properties. Using a DNA immunization strategy that includes intradermal (i.d.) immunization with naked plasmid DNA encoding the CD20 gene, we generated the hybridoma cell line D4, which secretes functional mAbs against an extracellular epitope of CD20. Immunocytochemistry analysis and a cell-based enzyme-linked immunosorbent assay using a Burkitt's lymphoma cell line showed that D4 mAbs are capable of binding to native extracellular epitopes of CD20. Moreover, the binding specificity of D4 mAbs was determined by western blot analysis. Cell proliferation was examined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was detected by the annexin V/propidium iodide staining and dye exclusion assay. The results showed that D4 anti-CD20 mAbs produced by DNA immunization exhibit potent growth inhibitory activity and have superior direct B-cell cytotoxicity compared to rituximab. We propose that antibody-induced apoptosis is one of the mechanisms of cell growth inhibition. Taken together, the data reported here open the path to DNA-based immunization for generating pharmacologically active monoclonal antibodies against CD20. In addition, the data support future in vivo animal testing and subsequent procedures to produce a potential therapeutic mAb.
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http://dx.doi.org/10.1016/j.intimp.2016.11.035DOI Listing
February 2017

Quantitative analysis of expression level of estrogen and progesterone receptors and VEGF genes in human endometrial stromal cells after treatment with nicotine.

Toxicol Mech Methods 2016 Oct 23;26(8):595-600. Epub 2016 Aug 23.

a Biochemistry Department, School of Medicine , Shiraz University of Medical Sciences , Shiraz , Iran.

Cigarette smoke is a complex mixture of toxic chemicals, including nicotine, carbon monoxide, and several recognized carcinogens and mutagens. Nicotine has a direct disturbing influence on steroid hormones (estrogen and progesterone), which are essential components of the female reproductive system, but the effect of nicotine on the hormone receptors is not yet clear. The aim of this study was to elucidate the effect of nicotine on the expression of estrogen receptor (ER), progesterone receptor (PR), and vascular endothelial growth factor (VEGF) in endometrial stromal cells. Expression levels of PR, ER, and VEGF in human endometrial stromal primary cells treated with nicotine (0, 10, 10, and 10μM) for 24 h were measured by quantitative real-time PCR. MTT assay demonstrated that nicotine decreased cell viability in a dose-dependent manner. Real-time PCR data showed that despite decrease in ER expression in the nicotine-treated groups compared with the control, nicotine exerted an increased inhibitory effect on PR expression compared to that on ER expression. VEGF mRNA expression in nicotine-treated endometrial stromal cells was increased. The results from this study provide novel evidence for inhibitory effects of nicotine on steroid hormones receptor expression in human primary endometrial cells. Also, our data suggest that nicotine might have angiogenesis effects on these cells.
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http://dx.doi.org/10.1080/15376516.2016.1218578DOI Listing
October 2016

Brain State-Dependent Transcranial Magnetic Closed-Loop Stimulation Controlled by Sensorimotor Desynchronization Induces Robust Increase of Corticospinal Excitability.

Brain Stimul 2016 May-Jun;9(3):415-424. Epub 2016 Feb 16.

Division of Functional and Restorative Neurosurgery, and Centre for Integrative Neuroscience, Eberhard Karls University, Tuebingen, Germany. Electronic address:

Background: Desynchronization of sensorimotor rhythmic activity increases instantaneous corticospinal excitability, as indexed by amplitudes of motor-evoked potentials (MEP) elicited by transcranial magnetic stimulation (TMS). The accumulative effect of cortical stimulation in conjunction with sensorimotor desynchronization is, however, unclear.

Objective: The aim of this study was to investigate the effects of repetitive pairing event-related desynchronization (ERD) with TMS of the precentral gyrus on corticospinal excitability.

Methods: Closed-loop single-pulse TMS was controlled by beta-band (16-22 Hz) ERD during motor-imagery of finger extension and applied within a brain-computer interface environment in eleven healthy subjects. The same number and pattern of stimuli were applied in a control group of eleven subjects during rest, i.e. independent of ERD. To probe for plasticity resistant to depotentiation, stimulation protocols were followed by a depotentiation task.

Results: Brain state-dependent application of approximately 300 TMS pulses during beta-ERD resulted in a significant increase of corticospinal excitability. By contrast, the identical stimulation pattern applied independent of beta-ERD in the control experiment resulted in a decrease of corticospinal excitability. These effects persisted beyond the period of stimulation and the depotentiation task.

Conclusion: These results could be instrumental in developing new therapeutic approaches such as the application of closed-loop stimulation in the context of neurorehabilitation.
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http://dx.doi.org/10.1016/j.brs.2016.02.007DOI Listing
September 2017

Efficient Conjugation of Aflatoxin M1 With Bovine Serum Albumin through Aflatoxin M1-(O-carboxymethyl) Oxime and Production of Anti-aflatoxin M1 Antibodies.

Jundishapur J Microbiol 2015 Apr 18;8(4):e16850. Epub 2015 Apr 18.

Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, IR Iran.

Background: Aflatoxins are the most extensively studied group of mycotoxins produced by molds, especially the Aspergillus group, which are highly toxic to animals and humans.

Objectives: Since immunoassay is a simple and rapid method for the analysis of many toxic substances in comparison to the chromatographic methods, it is necessary to produce specific and sensitive antibodies for detection of Aflatoxin M1 (AFM1). The current study was conducted to produce bioconjugate of Aflatoxin M1 (AFM1) with Bovine Serum Albumin (BSA) as well as to generate specific antibodies against AFM1 for immunoassay of the mycotoxin.

Materials And Methods: First, AFM1 was converted to AFM1-(O-carboxymethyl) oxime derivative. Then, AFM1-oxime was coupled with BSA and the product was assessed by UV-VIS spectrophotometry. In order to generate polyclonal antibodies against AFM1, rabbits were immunized with BSA-AFM1 conjugate. Produced antibodies were purified using ion exchange chromatography and BSA-Sepharose 4B affinity chromatography. The titers and specificity of the produced antibodies were determined by Enzyme-Linked Immunosorbent Assay (ELISA).

Results: The results indicated that coupling of AFM1 with O-(Carboxymethyl) hydroxylamine hemihydrochloride was suitable and 12 moles of AFM1-oxime were successfully coupled to each mole of BSA. In addition, the titers and specificity of the prepared antibody were considerable compared to standard anti-AFM1 antibodies. The relative cross-reactivity of each toxin (relative to AFM1) with purified anti-AFM1 antibodies, as determined by the amount of aflatoxin necessary to cause 50% inhibition of enzyme activity, was 70, 105, 240, and 2500 ng/mL for AFB1, AFB2, AFG1, and AFG2, respectively.

Conclusions: The prepared antibody can be used for the development of an ELISA kit to assay AFM1 in milk and other biological fluids.
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http://dx.doi.org/10.5812/jjm.8(4)2015.16850DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4449894PMC
April 2015

Learned self-regulation of the lesioned brain with epidural electrocorticography.

Front Behav Neurosci 2014 9;8:429. Epub 2014 Dec 9.

Institute for Medical Psychology and Behavioural Neurobiology, Eberhard Karls University Tuebingen Tuebingen, Germany ; Ospedale San Camillo, IRCCS Venice, Italy ; DZD, Eberhard Karls University Tuebingen Tuebingen, Germany.

Introduction: Different techniques for neurofeedback of voluntary brain activations are currently being explored for clinical application in brain disorders. One of the most frequently used approaches is the self-regulation of oscillatory signals recorded with electroencephalography (EEG). Many patients are, however, unable to achieve sufficient voluntary control of brain activity. This could be due to the specific anatomical and physiological changes of the patient's brain after the lesion, as well as to methodological issues related to the technique chosen for recording brain signals.

Methods: A patient with an extended ischemic lesion of the cortex did not gain volitional control of sensorimotor oscillations when using a standard EEG-based approach. We provided him with neurofeedback of his brain activity from the epidural space by electrocorticography (ECoG).

Results: Ipsilesional epidural recordings of field potentials facilitated self-regulation of brain oscillations in an online closed-loop paradigm and allowed reliable neurofeedback training for a period of 4 weeks.

Conclusion: Epidural implants may decode and train brain activity even when the cortical physiology is distorted following severe brain injury. Such practice would allow for reinforcement learning of preserved neural networks and may well provide restorative tools for those patients who are severely afflicted.
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http://dx.doi.org/10.3389/fnbeh.2014.00429DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4260503PMC
December 2014

Altered oxidative status and integrin expression in cyclosporine A-treated oral epithelial cells.

Toxicol Mech Methods 2015 Feb 22;25(2):98-104. Epub 2015 Jan 22.

Department of Orthodontics, Orthodontic Research Center, School of Dentistry, Shiraz University of Medical Sciences , Shiraz , Iran .

Background And Objective: Cyclosporine A (CsA) is an immunosuppressive agent administered to transplant patients. A well-known reported oral side effect of CsA consumption is gingival overgrowth (GO). Changes in the expression of integrins occurring in the gingiva following CsA treatment have been reported but these reports are mainly concerned with the connective tissue of the gingiva. In this study we targeted the alterations in the oral epithelium using KB cells, an oral epithelial cell line.

Methods: Cultured oral epithelial cells were treated with increasing concentrations of CsA (0.1, 1 and 10 µg/mL) and the molecular changes involving antioxidant enzymes [glutathione peroxidase (GPx) and glutathione reductase (GR)] and the level of reactive oxygen species (ROS) were measured. Quantitative real-time PCR was used to assess the expression of selected integrins (α2, α5 and β1).

Results: At CsA concentration above 0.1 µg/mL GPx demonstrated an increase in activity while GR activity and the level of reduced glutathione were diminished (p < 0.05). α5 and β1 integrin were downregulated at all treatment concentrations of CsA while α2 integrin presented this effect at concentrations above 1 µg/mL (p < 0.05).

Conclusion: The results suggest a possible role for oxidative stress and the altered expression of integrins in the pathology of CsA-induced gingival overgrowth.
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http://dx.doi.org/10.3109/15376516.2014.990595DOI Listing
February 2015

The combined effect of furosemide and propranolol on GSH homeostasis in ACHN renal cells.

Toxicol Mech Methods 2014 Sep 10;24(6):412-6. Epub 2014 Jun 10.

Reproductive Biology Department, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences , Shiraz , Iran .

Objectives: Propranolol, a beta-adrenergic blocker, is used in the treatment of a large number of cardiovascular diseases such as hypertension and arrhythmias. Propranolol, in combination with furosemide, is used to treat hypertensive disorders although their side effect profile is not very obvious. In present study, the effects of the drugs furosemide and propranolol were in corporately investigated both on glutathione homeostasis and their antioxidant effect on ACHN cells.

Methods: The cytoxicities and antioxidant effects of these two clinically important drugs on human kidney cell lines were evaluated using MTT following by the determination of glutathione reductase (GR) and glutathione peroxidase (GPx) activities and measuring the level of reduced glutathione (GSH).

Results: Propranolol induced a significant cytotoxic effect at 100 µM, while furosemide was cytotoxic at doses of 250 and 1000 µg/ml. A slight increase in GPx and GR activities and GSH level was observed with propranolol and furosemide treatment alone, while the two drugs together caused a significant increase in GPx and GR activities (35% and 42%, respectively) and GSH content (35%) in ACHN cell lysates (p < 0.05).

Conclusions: Our results demonstrate that although high doses of furosemide and propranolol are cytotoxic, co-administration of low doses may improve the antioxidant defense in patients undergoing treatment with these two important drugs.
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http://dx.doi.org/10.3109/15376516.2014.926437DOI Listing
September 2014

Proteolytic activities of kiwifruit actinidin (Actinidia deliciosa cv. Hayward) on different fibrous and globular proteins: a comparative study of actinidin with papain.

Appl Biochem Biotechnol 2014 Apr 7;172(8):4025-37. Epub 2014 Mar 7.

Department of Immunology, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran.

Actinidin, a member of the papain-like family of cysteine proteases, is abundant in kiwifruit. To date, a few studies have been provided to investigate the proteolytic activity and substrate specificity of actinidin on native proteins. Herein, the proteolytic activity of actinidin was compared to papain on several different fibrous and globular proteins under neutral, acidic and basic conditions. The digested samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and densitometry to assess the proteolytic effect. Furthermore, the levels of free amino nitrogen (FAN) of the treated samples were determined using the ninhydrin colorimetric method. The findings showed that actinidin has no or limited proteolytic effect on globular proteins such as immunoglobulins including sheep IgG, rabbit IgG, chicken IgY and fish IgM, bovine serum albumin (BSA), lipid transfer protein (LTP), and whey proteins (α-lactalbumin and β-lactoglobulin) compared to papain. In contrast to globular proteins, actinidin could hydrolyze collagen and fibrinogen perfectly at neutral and mild basic pHs. Moreover, this enzyme could digest pure α-casein and major subunits of micellar casein especially in acidic pHs. Taken together, the data indicated that actinidin has narrow substrate specificity with the highest enzymatic activity for the collagen and fibrinogen substrates. The results describe the actinidin as a mild plant protease useful for many special applications such as cell isolation from different tissues and some food industries as a mixture formula with other relevant proteases.
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http://dx.doi.org/10.1007/s12010-014-0812-7DOI Listing
April 2014

In Vitro Analysis of CsA-Induced Hepatotoxicity in HepG2 Cell Line: Oxidative Stress and α2 and β1 Integrin Subunits Expression.

Hepat Mon 2013 13;13(8):e11447. Epub 2013 Aug 13.

Biochemistry Department, Medical School, Shiraz University of Medicinal Sciences, Shiraz, IR Iran ; Recombinant Protein Laboratory, School of Advanced Medicinal Sciences and Technologies, Shiraz University of Medicinal Sciences, Shiraz, IR Iran.

Background: Cyclosporine A (CsA)-induced hepatotoxicity could be due to a reduction in α2β1 integrin expression that may either be from the direct effect of CsA itself or from reactive oxygen species (ROS) overproduction.

Objectives: In this study we aimed to identify the cellular mechanisms underlying CsA-induced hepatic injury by investigating the activation patterns of the antioxidant enzymes, using HepG2 as an in vitro model.

Materials And Methods: HepG2 cells were cultured with different concentrations of CsA (0, 0.1, 1, 10 μg/ml) for 72 h. Effect of CsA on, 1) cellular integrity, 2) glutathione reductase (GR) and glutathione peroxidase (GPx) activity, 3) cellular levels of glutathione (GSH), 4) intracellular ROS, 5) ALT and AST activities, 6) urea production and 7) α2β1 integrin expression were assayed.

Results: CsA treatment demonstrated a dose dependent increase in intracellular levels of ROS, GPx activity and decrease in GSH levels (P<0.05). GR activity was mildly attenuated in 1 and 10 µg/ml concentrations of CsA. Alanine aminotranferase (ALT) and aspartate aminotransferase (AST) levels increased in CsA treated cells, while urea synthesis was significantly decreased following treatment with higher concentrations of CsA (P<0.05). Significant down-regulation of β1integrin expression was observed in 1 and 10 µg/ml CsA treated cells while α2 integrin mRNA was significantly down-regulated in all CsA treated cells.

Conclusions: The observed reduction of α2β1 integrin expression following CsA treatment could be proposed as a possible pathway of CsA-induced hepatotoxicity. Further studies are required to elucidate whether this attenuated expression is due to the direct effect of CsA or caused by overproduction of ROS.
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http://dx.doi.org/10.5812/hepatmon.11447DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3785933PMC
October 2013

An efficient method for purification of nonspecific lipid transfer protein-1 from rice seeds using kiwifruit actinidin proteolysis and ion exchange chromatography.

J Sep Sci 2012 Nov 17;35(21):2827-33. Epub 2012 Sep 17.

Department of Biology, Faculty of Sciences, Razi University, Kermanshah, Iran.

Plant nonspecific lipid transfer proteins are small basic proteins that transport phospholipids between membranes and are subdivided into two subfamilies, nsLTP(1) (9 kDa) and nsLTP(2) (7 kDa). LTPs have potential application in the defense reactions against pathogens and the drug delivery systems. Many efforts have been made for purification of different nsLTPs from various plants; however, most of them used successive purification procedures. We have developed a relatively simple and efficient method for the purification of rice nsLTP(1), based on the proteolytic activity of kiwifruit actinidin on the rice seed extract and one-step chromatographic procedure on a CM-Sepharose column. The purity of protein was determined by reversed-phase high-performance liquid chromatography (RP-HPLC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The isolated LTP(1) migrated as a homogenous polypeptide with molecular mass of 9 kDa that confirms the efficiency of actinidin on the digestion of major contaminations present in the rice seed extract without any harmful effect on the LTP(1). The advantages of using proteolytic activity of actinidin in purifying rice LTP(1) includes the reduced separation time allowing the purification of LTP(1) in one-step chromatographic procedure, low costing, high efficiency, and the relative simplicity of the method.
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http://dx.doi.org/10.1002/jssc.201200383DOI Listing
November 2012

Effect of urea on protein separation by ion-exchange chromatography.

J Biochem 2010 May 4;147(5):735-41. Epub 2010 Feb 4.

Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran.

Ion-exchange chromatography (IEC) is the most frequently used chromatographic technique for the separation of proteins and peptides. In this article, the effects of urea on IEC separation of kiwifruit actinidin, egg white and urinary proteins were examined. The purity and relative amount of each protein in different conditions (in the presence or absence of urea) were compared with each other. The three parameters, including resolution, selectivity and efficiency of column in the presence of urea, were calculated and compared with the absence of urea. The results revealed that urea improved the purity of proteins and the resolution, selectivity and efficiency of IEC in separation of studied proteins.
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http://dx.doi.org/10.1093/jb/mvq008DOI Listing
May 2010

Purification of three major forms of beta-hCG from urine and production of polyclonal antibodies against them.

Clin Biochem 2009 Sep 6;42(13-14):1476-82. Epub 2009 Jun 6.

Department of Biochemistry, Faculty of Science, Payame Noor University, Tehran, Iran.

Objectives: This study has been conducted to develop a proper and relatively simple method for purification of three major forms of beta-hCG from urine to use as clinical calibrators and also to produce anti-beta-hCG polyclonal antibodies against these purified forms.

Design And Methods: Major beta-hCG forms from urine of pregnant women were purified using an ultrafiltration method followed by three successive steps: 1) affinity chromatography on a Concanavalin A-Sepharose column, 2) ion exchange chromatography on a DEAE-Sephacel column and 3) preparative gel electrophoresis. Specific polyclonal antibodies against the purified forms were produced by immunizing rats. We also extracted beta-hCG from urine using trypsin affinity chromatography.

Results: Three beta-hCG forms with the molecular mass (MW) of 28, 32 and 35 kDa were purified and identified by ELISA method using monoclonal antibodies against two distinct epitopes (beta2 and beta11) on beta-hCG. The titer of prepared antibodies was comparable with common standard anti-beta-hCG antibodies. Also we could purify a 37 kDa form of beta-hCG by trypsin affinity chromatography.

Conclusions: Here we proposed two different proper methods for purification of major forms of beta-hCG from urine. We also found that three prepared beta-hCG forms expose the beta11 epitope and have an hCGbetacf epitope expressed. Specific antibodies against purified beta-hCG forms had comparable titers and could be used in diagnostic kits.
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http://dx.doi.org/10.1016/j.clinbiochem.2009.05.019DOI Listing
September 2009