zakaria vahabzadeh | Kerman Medical University (original) (raw)

Papers by zakaria vahabzadeh

Clinica chimica acta, May 1, 2024

Biochemical genetics, Mar 27, 2024

Gene Reports, Feb 29, 2024

Advances in Medical Sciences, Mar 1, 2018

Trimethylamine N-oxide (TMAO) is a biomarker for kidney problems. It has also been introduced as ... more Trimethylamine N-oxide (TMAO) is a biomarker for kidney problems. It has also been introduced as a risk factor for atherosclerosis. The classic risk factors for atherosclerosis trigger cellular and humeral immunoreaction in macrophages through induction of heat shock protein expressions and increased levels of GRP94 and HSP70 are associated with increased atherosclerosis risk. The present study evaluated the possible effect(s) of TMAO on the expression of GRP94 and HSP70 at protein levels. Methods: J774A.1 murine macrophages were treated with different micromolar concentrations of TMAO and 4-phenylbutyric acid (PBA), a chemical chaperone, for 8, 18, 24, and 48 h intervals. Tunicamycin was also used as a control for induction of endoplasmic reticulum stress. Western blotting was used to evaluate the expression of GRP94 and HSP70 in macrophages at protein levels. Result: Tunicamycin greatly increased protein levels of GRP94. Similarly, but to a lesser extent compared to tunicamycin, TMAO also increased GRP94. In 24 h treated cells, only 300 mM of TMAO, and in cells treated for 48 h, all doses of TMAO produced a significant increase in relative HSP70 protein levels compared to the control. PBA failed to induce any changes in GRP94 or HSP70 protein levels. Conclusion: GRP94 and HSP70 are stress-inducible heat shock protein, so the elevation in J774A.1 murine macrophages can clearly define cells under stress and elucidate the contribution of stress induced by TMAO that may have a part in the abnormal activation of macrophages involved in foam cell formation.

PubMed, Jul 1, 2021

Background: Trimethylamine N-oxide (TMAO) is emerging as a new generation of metabolites related ... more Background: Trimethylamine N-oxide (TMAO) is emerging as a new generation of metabolites related to the activation of inflammatory reactions in the macrophages during atherosclerosis. Stress-activation of cell surface toll-like receptors (TLRs) as well as nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (NOX) is also assumed to be involved in TMAO-induced inflammatory reaction in the macrophages. To elucidate the possible contribution of TLRs and NOX to the mentioned signaling pathway, we aimed to simultaneously evaluate the expression level of TLR2, TLR6, and NOX2 in TMAO-treated macrophages. Methods: 2.5 × 106 cells of U937-derived macrophages were treated in triplicates with different concentrations (37.5, 75, 150, and 300 μM) of TMAO for 24 hours. The cells were also treated with tunicamycin (TUN), as a positive control of stress. Normal control group (CTR) cells received no treatment. The viability of treated cells was checked by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a tetrazole (MTT) assay. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was also used to evaluate the relative expression (fold change) of TLR2, TLR6, and NOX2 at messenger ribonucleic acid (mRNA) levels. One-way analysis of variance (ANOVA) with post-hoc Dunnett's test was performed to compare every mean with that of the control. Results: No cell death occurred because of treatments. Dose of 300 μM of TMAO significantly increased the relative expression of both TLR2 and NOX2 compared to the CTR cells (P < 0.001 for both). The elevation of TLR6 was not statistically significant in all groups of TMAO-treated cells (P > 0.050). Conclusion: Our results provide documentation supporting contribution of TLR2 and NOX2 to previously described inflammatory reactions induced by TMAO in macrophages. In addition, they may clarify the proatherogenic role of TMAO in foam cell formation as well as abnormal activation of macrophages during atherosclerosis.

PubMed, Sep 1, 2017

Introduction: Hyperglycemia is a common occurrence in critically ill patients, and its prevalence... more Introduction: Hyperglycemia is a common occurrence in critically ill patients, and its prevalence in patients receiving nutritional support is much higher than in other patients. The non-diabetic form is associated with more undesirable outcomes. This study was performed to determine the prevalence of non-diabetic hyperglycemia and its correlates in patients receiving enteral nutrition. Material and methods: This cross-sectional study was performed between March and December 2015. Seven hundred forty eight (748) patients were reviewed to see if they met the inclusion criteria. After random sequence numbering, 414 patients who were eligible for further assessment and data gathering were selected. Hyperglycemia was defined as the blood glucose levels higher than either 126 mg/dL, in the fasting state, or 180 mg/dL, in a random state. Blood glucose was measured by an ACCU-CHECK glucometer (Roche diagnostics, Mannheim, Germany) three times, after ICU admission, in both fasting and random state. A pre-prepared form was used to extract data from hospital records. Data analysis was performed by SPSS 21 software. Results: In this group of hospitalized patients, the prevalence of non-diabetic hyperglycemia was 14/49 (60/414). In the hyperglycemic subgroup, mean FBS was 228.00±36.42, mean random BS was 183.19±43.94 and mean blood sugar on the first day of hospitalization was 203.60 ± 60.79. The mean age of patients was 56.64±19.79 years and the mean duration of hospitalization was 19.24±15.33 days. There was no significant relationship between enteral nutrition feeding volume and hyperglycemia. Majorly, patients aged above 60 years were hyperglycemic. The prevalence was higher in men than in women. Most patients were internal cases, but with the highest prevalence of hyperglycemia in surgical patients. Conclusions: Since among different studied variables just diagnosed disease and the level of provided calorie showed significant differences between subgroup categories, so it can be suggested that designing on-time appropriate management programs based them can be effective on the administration of non-diabetic hyperglycemia and its undesirable consequences in such patients.

PubMed, Feb 6, 2021

Purpose: The aim of the present study is to assess the effect of L-carnitine and Coenzyme Q10 (Co... more Purpose: The aim of the present study is to assess the effect of L-carnitine and Coenzyme Q10 (CoQ10) on human sperm motility, DNA fragmentation, chromatin structure, and reactive oxygen species (ROS) during, before and after freezing in oligospermia men. Materials and methods: Semen was collected from 30 oligospermic men, who referred to infertility clinic of Beasat Hospital in Sanandaj, Iran. The samples of each individual were divided into 8 equal parts: 1. control group before freezing; 2. incubated with L-carnitine; 3. incubated with coenzyme Q10; 4. incubated with the combination of L-carnitine + CoQ10; 5. control freezing group; 6. the experimental freezing group with L-carnitine; 7. the experimental freezing group with coenzyme Q10 and 8. the experimental freezing with the combination of L-c + CoQ10. Sperm motility was assessed by WET MOUNT method. DNA fragmentation was evaluated by SCD (Sperm Chromatin Desperation), ROS, was evaluated by quantitative fluorescence reaction, and chromatin deficiency was determined by chromatin staining (CMA3). Results: Antioxidant treatments, significantly reduced the number of ROS + in the pre and post freezing groups. Significant improvement was seen in the sperm motility of class B in the pre freezing groups with L-carnitine. Antioxidants also reduced the percentage of DNA fragmentation and protamine deficiency in pre-and post-freezing. Conclusion: Addition of Coq10 and L-carnitine to human sperm medium significantly reduced the number of ROS. This reduction in ROS reduced sperm damage during cryopreservation.

Journal of Cellular Biochemistry, Feb 3, 2019

Family with sequence similarity 83 member H (FAM83H) protein‐coding geneplay an essential role in... more Family with sequence similarity 83 member H (FAM83H) protein‐coding geneplay an essential role in the structural organization, calcification of developing enamel, and keratin cytoskeleton disassembly by recruiting Casein kinase 1 alpha (CSNK1A1) to keratin filaments. In this study, we have applied CRISPR Cas9 nickase (D10A) to knockout (KO) the Fam83h gene in NMRI outbred mice. We generated homozygous Fam83h KO mice ( Fam83h Ko/Ko) through a premature termination codon, which was validated by Sanger sequencing in F0 generation. Next, we also bred the FAM83H KO for two generations. Reverse‐transcription polymerase chain reaction and Western blot analysis approved the Fam83h KO mice. The Fam83h KO mice had evidence of normal morphology at the cervical loops, secretory and maturation stages, and mandibular molars. In comparison with the normal wild‐type mice ( Fam83h W/W), the F2 homozygous KO ( Fam83h Ko/Ko) had sparse, scruffy coats with small body size and decreased general activity. Also, they had the natural reproductive ability and natural lifespan. In addition, delay in opening the eyes and dry eyes among infant mice were seen. The F1 heterozygous mice looked comparable to the normal wild‐type mice ( Fam83h W/W), which showed autosomal recessive inheritance of these phenotypes. The KO of FAM83H had controversial effects on the development of teeth and the formation of enamel. The phenotype defect in dental development and the enamel formation were seen in three mice among four generations. It can be concluded that null FAM83H in outbred mice not only showed the reported phenotypes in null inbred mouse but also showed normal lifespan and reproductive ability; dental deficiency in three homozygous mice; and the symptoms that were similar to the symptoms of dry eye syndrome and curly coat dog syndrome in all four evaluated KO generations.

Research Square (Research Square), May 13, 2022

The authors have requested that this preprint be removed from Research Square.

Neurochemical Research, Mar 1, 2023

Journal of Physiology and Biochemistry, Jan 10, 2022

Nonalcoholic fatty liver disease (NAFLD) is a common chronic liver disease that is rapidly becomi... more Nonalcoholic fatty liver disease (NAFLD) is a common chronic liver disease that is rapidly becoming a public health problem. An imbalance in lipid distribution to the hepatocytes and metabolism causes hepatocyte steatosis. Vaspin is a newly discovered adipokine that has been linked to a variety of metabolic disorders. The effects of vaspin on steatosis and fibrosis pathogenesis and related mechanisms are unclear. Thus, this study investigated the molecular mechanism of vaspin on hepatocyte steatosis and fibrosis. HepG2 cells were treated with 1.2 mM free fatty acid and the intracellular lipid values were measured by flow cytometry and Nile red assay. RT-qPCR was used to assess the effect of vaspin and blocking of the GRP78 receptor on the expression of lipogenesis, oxidation, uptake, and secretion of fatty acid (FA), as well as AMPK activity. In co-cultured HepG2 and LX-2 cell lines, the expression of main proteins of hepatocyte fibrosis was analyzed using Western blot analysis. In the HepG2 cell line, we discovered that vaspin increased oxidation, FA secretion and gene expression, and AMPK activity and decreased lipogenesis and FA uptake and gene expression. Western blot analysis in co-cultured HepG2 and LX-2 cell lines showed that α-SMA and TGF-β1 protein expression decreased. The data demonstrated that vaspin acts as a novel regulator of hepatocyte steatosis through the GRP78 receptor, effectively reducing hepatocyte fibrosis through AMPK activation and decreasing NF-κB gene expression.

Molecular Biology Reports, Mar 29, 2022

BACKGROUND Non-alcoholic fatty liver disease (NAFLD) is a multifactorial disorder with complicate... more BACKGROUND Non-alcoholic fatty liver disease (NAFLD) is a multifactorial disorder with complicated pathophysiology. Trimethylamine-N-oxide (TMAO) has been thought to be correlated with the pathogenesis of NAFLD. The single nucleotide polymorphisms (SNPs) of hepatic flavin-containing monooxygenase 3 (FMO3) regulate the concentration of TMAO. This case-control study investigated the plasma levels of TMAO as well as its possible correlation with the frequency of specific genotype of FMO3 (-2650C>G, -2543T>A, -2177G>C, -2589C>T, -2106G>A polymorphisms) in Kurdish patients with NAFLD. METHODS AND RESULTS: In 85 confirmed NAFLD patients and 30 healthy individuals, triglycerides (TG), total cholesterol (Chol), low-density lipoprotein (LDL), high-density lipoprotein (HDL), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activities were measured. TMAO was also measured using the LC-MS/MS method. High-resolution melting analysis was applied to determine FMO3 genotypes. Plasma TMAO levels were significantly higher in patients (p = 0.030). A CC genotype with a frequency of 12.9% for SNP -2177G>C was found in Kurdish NAFLD patients. The distribution of the GC genotype was also significantly different (p = 0.017). CONCLUSIONS The current results provide documentation for high circulatory levels of TMAO and its possible correlation with the presence of the specific genotype -2177G>C FMO3 in Kurdish NAFLD patients.

Scientific journal of Kurdistan University of Medical Sciences, May 1, 2022

Background and Aim: Non-alcoholic fatty liver disease, characterized by abnormal fat accumulation... more Background and Aim: Non-alcoholic fatty liver disease, characterized by abnormal fat accumulation in the liver, is associated with obesity and insulin resistance. Vaspin is an adipokine secreted by adipose tissue, and its gene expression increases when insulin sensitivity is reduced. In this study, we made a comparison between 3D and 2D cultures in regard to the effects of vaspin on fatty acid metabolism. Materials and Methods: The steatosis model was induced by oleic and palmitic acid in HepG2 cell line in two-and three-dimensional cultures (collagen gel). Then, the cells were treated with 100 ng/ml vaspin for 24 hours. We used Real time PCR for measurement of the expressions of FABP4, MCAD, FAS and ApoB100 genes. Results: In two-and three-dimensional cultures, we found significant decrease in the expression of FABP4 and FAS genes. There was no significant difference between the expression of these genes in the two-and three-dimensional cultures. We detected significant increase in the expression of MCAD and ApoB100 genes in 2D and 3D cultures. We did not find any significant difference in the expressions of MCAD and ApoB100 genes between two-and three-dimensional cultures. Conclusion: Alteration of the expressions of the genes involved in uptake, lipogenesis, oxidation and secretion of fatty acids occurred in the group treated with vaspin compared to the control group in the 2-D and 3-D cultures. But, we did not find any significant difference in the expressions of these genes between the 2-D and 3-D cultures.

Zygote, Apr 29, 2022

SummaryMouse embryonic fibroblast (MEF) cells are commonly used as feeder cells to maintain the p... more SummaryMouse embryonic fibroblast (MEF) cells are commonly used as feeder cells to maintain the pluripotent state of stem cells. MEFs produce growth factors and provide adhesion molecules and extracellular matrix (ECM) compounds for cellular binding. In the present study, we compared the expression levels of Fgf2, Bmp4, ActivinA, Lif and Tgfb1 genes at the mRNA level and the level of Fgf2 protein secretion and Lif cytokine secretion at passages one, three and five of MEFs isolated from 13.5-day-old and 15.5-day-old embryos of NMRI and C57BL/6 mice using real-time PCR and enzyme-linked immunosorbent assay. We observed differences in the expression levels of the studied genes and secretion of the two growth factors in the three passages of MEFs isolated from 13.5-day-old and 15.5-day-old embryos, respectively. These differences were also observed between the NMRI and C57BL/6 strains. The results of this study suggested that researchers should use mice embryos that have different genetic backgrounds and ages, in addition to different MEF passages, when producing MEFs based on the application and type of their study.

Although there is little evidence that modest amounts of fructose have detrimental effects on car... more Although there is little evidence that modest amounts of fructose have detrimental effects on carbohydrate and lipid metabolisms, larger doses of fructose have been associated with numerous metabolic abnormalities and induce oxidative stress in human and laboratory animals. This study was done to investigate the effects of high fructose enriched diet (65.1%) on plasma lipid profile and some markers of oxidative stress in heart and kidney in rats. Materials & Methods: Sixteen Male adult Wistar rats with body weight of 213±5 g were divided into two control and Fructose groups each group consisted of eight rats which received their appropriate diets for a six-week period. Food and fluid intake thrice and body weight changes were monitored weekly. At the end of the experimental period, plasma was collected and used for measurement of HDL-C, LDL-C, total cholesterol, triglyceride (TG) and total antioxidant capacity by appropriate kits. The heart and kidneys were homogenized in phosphate-buffered saline (pH7.4) and in their Supernatants Glutathione peroxidase (GPX), superoxide dismutase (SOD) and malondialdehayde (MDA) were measured by appropriate kits. Independent sample T-test was used for statistical analysis in SPSS. Results: Fructose made no significant difference in body weight, fluid intake, and total antioxidant capacity (P>0.05). Fructose significantly decreased food intake of rats at the end of experimental period (P=0.000). Fructose significantly increased TG (P=0.040) in plasma and MDA in heart (p=0.000) and kidneys (P=0.002), reduced activity of GPX in heart (P=0.052) and kidneys (P=0.018) and SOD in heart (P=0.047) and kidneys (P=0.034) of rats compare with the control group. Conclusion: This study shows that Fructose enriched diet significantly increases TG in plasma, MDA in heart and kidneys and reduces GPX and SOD in heart and kidneys of rats. Using this diet may enhance metabolic dysfunctions and oxidative stress in rats.

PubMed, Sep 26, 2015

Trimethylamine N-oxide (TMAO), a common metabolite in animals and humans, can induce changes in t... more Trimethylamine N-oxide (TMAO), a common metabolite in animals and humans, can induce changes in the expression or conformation of heat shock proteins. It has also been introduced as a risk factor for atherosclerosis and a biomarker for kidney problems. On the other hand, increased levels of heat shock proteins 60 and 70 KDa are associated with increased atherosclerosis risk. This study was therefore designed to evaluate the possible effect(s) of TMAO on the expression of HSP60 and GRP78 at the mRNA and protein levels. Murine macrophage J774A.1 cells were treated with micromolar concentrations of TMAO and 4-phenylbutyric acid (4-PBA), a chemical chaperon, for different time intervals. Tunicamycin was also used as a control for induction of endoplasmic reticulum stress. Tunicamycin greatly increased both mRNA and protein levels of GRP78. Similarly but to a lesser extent compared to tunicamycin, TMAO also increased mRNA and protein levels of GRP78 in a dose and time-dependent manner. In contrast, 4-PBA failed to induce any changes. Similar to GRP78, HSP60 was also increased only at mRNA level in TMAO treated cells. 4-PBA also increased HSP60 mRNA levels, whereas, tunicamycin did not show any effect on either protein or mRNA levels of HSP60. Since both heat shock proteins are stress inducible and the elevation of GRP78 is a hallmark for endoplasmic reticulum stress induction, it can be concluded that TMAO may induce endoplasmic reticulum stress or may act through elevation of these heat shock proteins.

Current Nutrition & Food Science

Aims: The aim of the present study was to assess and compare the effect of different methods of c... more Aims: The aim of the present study was to assess and compare the effect of different methods of cooking on B-group vitamins (B1, B2, and B6), As, Cd, Ni, Pb, and some essential elements (Cu, Co, and Mn) in the main local varieties of Iranian rice. Background: Different cooking procedures have been shown to have contradictory benefits in terms of nutritional value by affecting the bioavailability of vitamins and microminerals. Objective: The aim of the present study was to assess and compare the effect of different methods of cooking on B-group vitamins (B1, B2 and B6), As, Cd, Ni, Pb, and some essential elements (Cu, Co, and Mn) in the main local varieties of Iranian rice. Method: The effectiveness of different cooking procedures, including direct cooking (Dampokht) and cooking by washing the rice with water and draining off the water (Abkesh), on B1, B2, and B6 vitamins contents and Cd, As, Pb, Ni, Cu, Co, Mn elements in the different local varieties of Iranian rice was evaluated. ...

Research Journal of …, 2009

Abstract: Simultaneous effects of processing factors such as ripening time (20-60 days), ripening... more Abstract: Simultaneous effects of processing factors such as ripening time (20-60 days), ripening temperature (5-10°C), level of rennet added (1-2 g/100 kg milk), brine concentration (12-16% wv -1 ), type of brine (NaCl, 25% Kcl + 75% NaCl) and level of starter (1, 3%) on biogenic ...

PubMed, Feb 1, 2024

Objective: There is interest in using cytotoxic T lymphocyte antigen-4 (CTLA-4) immunotherapy to ... more Objective: There is interest in using cytotoxic T lymphocyte antigen-4 (CTLA-4) immunotherapy to treat blood cancers. Unfortunately, patients with acute lymphoblastic leukaemia (ALL) frequently exhibit resistance to treatment and natural killer (NK) cell exhaustion. This study aims to increase the cytotoxic potency of natural killer cells by using CTLA-4 to block the Nalm-6 leukaemia cell line. Materials and methods: In this experimental study, NK cells were purified from the peripheral blood mononuclear cells (PBMCs) of 10 healthy people and assessed by flow cytometry for purity and viability. The purified cells were activated overnight at 37°C and 5% CO2 with interleukin-15 (IL-15, 10 ng/ml) followed by evaluation of expressions of CTLA-4, activating and inhibitory receptors, and the release of interferon gamma (IFN-γ) and granzyme B (GZM B). CTLA-4 expression on NK cells from recurrent ALL patients was also evaluated. Finally, the cytotoxic activity of NK cells was assessed after the CTLA-4 blockade. Results: The purity of the isolated cells was 96.58 ± 2.57%. Isolated NK cells activated with IL-15 resulted in significantly higher CTLA-4 expression (8.75%, P<0.05). Similarly, CTLA-4 expression on the surface of NK cells from patients with ALL was higher (7.46%) compared to healthy individuals (1.46%, P<0.05). IL-15 reduced NKG2A expression (P<0.01), and increased expressions of NKP30 (P<0.05) and NKP46 (P<0.01). The activated NK cells released more IFN-γ (P<0.5) and GZM B (P<0.01) compared to unactivated NK cells. Blockade of CTLA-4 enhanced the NK cell killing potential against Nalm-6 cells (56.3%, P<0.05); however, IFN-γ and GZM B levels were not statistically different between the blocked and non-blocked groups. Conclusion: Our findings suggest that CTLA-4 blockage of Nalm-6 cells causes an increase in antitumour activity of NK cells against these cells. Our study also provides evidence for the potential of cancer immunotherapy treatment using blocking anti-CTLA-4 mAbs.

Biosciences, Biotechnology Research Asia, Sep 25, 2016

To investigate the effect of lecithin on FMO3 and FMO5 gene expression, L-α α α α αlecithin or et... more To investigate the effect of lecithin on FMO3 and FMO5 gene expression, L-α α α α αlecithin or ethanol 30% (as control) was administrated to female N-mary mice as intraperitoneal injection and animals were sacrificed 48h after treatment. The relative expression of mRNA and protein were assessed by real-time PCR and western blotting analysis. In this study, FMO5 expression was also evaluated in HepG2 cell line. FMO3 mRNA expression in mice liver was increased to 1.8-fold in animal received lecithin compared to control group, While FMO5 expression did not showed any changes as in vivo and in vitro experiments. The our results suggest to induce of FMO3 expression by lecithin.