zakaria vahabzadeh

Huntington's disease is a chronic hereditary disorder that causes cognitive and movement defects in affected individuals by progressive destruction of neurons in the cerebral cortex, striatum and the hippocampus. Studies have shown that increased activity of cyclin-dependent kinase-5 (CDK5) plays an important role in the pathogenesis and occurrence of memory impairment in Huntington's disease. Recently, alpha-pinene has been reported to improve learning and memory performance in Alzheimer's and Parkinson's models. Therefore, the aim of this study was to investigate the effect of alpha-pinene on passive avoidance memory and CDK5 gene expression in Huntington's animal model induced by 3-nitro-propionic acid (3-NP).

In this study, 40 male rats were randomly divided into 5 groups: sham, 3-NP (10 mg/kg),and 3 other groups receiving 3-NP (10 mg/kg) + alpha-pinene at doses of 1, 5 and 10 mg/kg (for 3 weeks,via intraperitoneal injection). Passive avoidance memory was assessed through the shuttle box device. Then, the expression level of CDK5 gene was measured by RT-qPCR method in brain cortex and hippocampus.

3-NP injection caused memory impairment by decreasing step through latency (STL). Alpha-pinene at all three doses improved passive avoidance memory performance. Also, 3-NP injection caused a significant increase in CDK5 gene expression level in the brain cortex and hippocampus compared to that in the sham group. The groups which received alpha-pinene at doses of 5 and 10 mg/kg in brain cortex and 1 mg/kg in hippocampus showed decreased expression level of this gene compared to the group that received 3-NP.

The results of this study showed that alpha-pinene improves passive avoidance memory performance probably by reducing the CDK5 gene expression level in Huntington's animal model induced by 3-NP.

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.

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.

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.

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.

Dental enamel formation is a complex process that is regulated by various genes. One such gene, Family With Sequence Similarity 83 Member H (Fam83h), has been identified as an essential factor for dental enamel formation. Additionally, Fam83h has been found to be potentially linked to the Wnt/β-catenin pathway.

This study aimed to investigate the effects of the Fam83h knockout gene on mineralization and formation of teeth, along with mediators of the Wnt/β-catenin pathway as a development aspect in mice.

To confirm the Fam83h-KnockOut mice, both Sanger sequencing and Western blot methods were used. then used qPCR to measure the expression levels of genes related to tooth mineralization and formation of dental root, including Fam20a, Dspp, Dmp1, Enam, Ambn, Sppl2a, Mmp20, and Wnt/β-catenin pathway mediators, in both the Fam83h-Knockout and wild-type mice at 5, 11 and 18 days of age. also the expression level of Fgf10 and mediators of the Wnt/β-catenin pathway was measured in the skin of both Knockout and wild-type mice using qPCR. A histological assessment was then performed to further investigate the results.

A significant reduction in the expression levels of Ambn, Mmp20, Dspp, and Fgf10 in the dental root of Fam83h-Knockout mice compared to their wild-type counterparts was demonstrated by our results, indicating potential disruptions in tooth development. Significant down-regulation of CK1a, CK1e, and β-catenin in the dental root of Fam83h-Knockout mice was associated with a reduction in mineralization and formation-related gene. Additionally, the skin analysis of Fam83h-Knockout mice revealed reduced levels of Fgf10, CK1a, CK1e, and β-catenin. Further histological assessment confirmed that the concurrent reduction of Fgf10 expression level and Wnt/β-catenin genes were associated with alterations in hair follicle maturation.

The concurrent reduction in the expression level of both Wnt/β-catenin mediators and mineralization-related genes, resulting in the disruption of dental mineralization and formation, was caused by the deficiency of Fam83h. Our findings suggest a cumulative effect and multi-factorial interplay between Fam83h, Wnt/Β-Catenin signaling, and dental mineralization-related genes subsequently, during the dental formation process.

Oxidative stress is an important factor in the development of memory and learning disorder which can cause neuronal damage in the hippocampus. Alpha-pinene is a polyphenolic compound from the terpene family that has shown important anti-inflammatory, anti-anxiety, antioxidant and neuroprotective effects in the central nervous system and can affect memory. The aim of the present study was to investigate the effect of alpha-pinene on the improvement of working and spatial memory in rats. 

In this study, 24 male rats were randomly divided into 3 groups: control and 2 alpha-pinene groups (5 and 10 mg/kg IP) for 3 weeks. Spatial and working memories were assessed by Morris water maze and Y maze, respectively. Then, malondialdehyde level and total antioxidant capacity in hippocampal tissue were measured. Data were analyzed using one-way analysis of variance and Tukey's post hoc test.

The percentage of alternation in the Y maze increased in the group which had received 10 mg/kg alpha-pinene group compared to those in the control group and the group which had received 5 mg/kg alpha-pinene. The time spent in the target area at the dose of 10 mg/kg of alpha-pinene showed a significant increase compared to that in the control group, but there was no significant difference among the groups in terms of the time to reach the target platform. Alpha-pinene at the dose of 10 mg/kg decreased the level of malondialdehyde in hippocampal tissue compared to the control group, but no significant difference was observed between the groups in terms of total antioxidant capacity.

Alpha-pinene increased spatial and working memory performance in rats. One of the possible mechanisms of memory improvement in the present study could be due to the reduction of malondialdehyde in the hippocampal tissue, as one of the important indicators of oxidative stress in the central nervous system.

Fam83h Protein is a non-secretory protein that interacts with Casein Kinase1Alpha1(CSNK1A1) through its N-terminal.

  In this study, the C-terminal domains of Fam83h in human and mouse were modeled using the I-TASER software for prediction of protein structure and function. The molecular dynamics (MD) simulations were performed by GROMACS version 5.0.2 to investigate their temporal behavior. FEL analysis was done for each model after MD. Protein-protein docking was done by PIPER to investigate the interaction of Fam83h C-terminal with cytoskeletal keratins5 as a cellular keratin model.

The results showed that the human protein is more stable and compact than the mouse protein. Assessment of the interaction between the C-terminal of the mouse Fam83h and Keratin-5 proteins showed the residues Arg378, Pro391, Ser663, Glu710, Arg923 in mouse Fam83h protein made hydrogen bonds with Leu474, Arg417, Tyr453, Glu 397, Gln 396 and Glu 390 of the chain A and B of mouse keratin-5, respectively. Human Fam83h and keratin 5 form hydrogen bonds via residues of Thr433, His447 and Arg477 and Leu473, Gly476, Glu420 and Arg407.

According to the previous experimental reports, Fam83h can interact with keratin cytoskeleton and has a role in cancer progression. It can be concluded that Fam83h protein can directly interact with keratin filaments through its C-terminal. Therefore, our hypothesis based on  in-silico study is: non-sense mutation in C-terminal of Fam83h protein may lead to truncated protein and subsequently disruption of keratin cytoskeleton bundling which can be regarded as a mechanism involved in cancer progression.

Propolis, as a natural product have several biological benefits including antioxidant, anti-inflammatory, and antimicrobial as well as anticancer properties. Its application extensively depends on its composition and the concentrations of the major active components. The aim of, this study was to evaluate the concentrations of chrysin, quercetin and, caffeic acid as three active components, in the Iranian propolis using UV-HPLC method.

Three different samples of propolis were collected from Saral and Avalan regions (Kurdistan Province, Iran) in spring 2017. To separate and measure the concentrations of the three components, the samples were extracted by using an ethanolic solution (80%) and subsequently analyzed by the UV-HPLC system with an isocratic mode at a flow rate of 1ml/min for 20 min and ultraviolet detection at λmax 260 or 310 nm. For evaluation of the quality control of the assay, the linearity and detection limits of the method were calculated.

Chrysin was not detected in our samples. The average concentrations of quercetin and caffeic acid were 65.425 and 27.498 µg/g, respectively.

For the first time, we reported accurately the concentrations of three active components of Iranian propolis which many of propolis's beneficial effects have been attributed to them.

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.

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.

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.

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.

Trimethylamine-N-Oxide (TMAO) is considered as a risk factor for atherosclerosis which further leads to inflammation during atherosclerosis. The exact mechanism(s) by which TMAO induces the inflammatory reactions remains to be determined. TMAO can cause the endoplasmic reticulum (ER) stress that triggers activation of Toll-Like Receptors (TLRs). In macrophages, this process stimulates the production of proinflammatory cytokines. This study designed to evaluate the expression level of TLR4 in TMAO-treated macrophages.

In this experimental study, different concentrations of TMAO (37.5, 75, 150, and 300 μM) were exposed to murine macrophage (J774A.1 cell line) for 8, 18, 24, and 48 hours. The cells were also treated with 2.5 mM of 4-phenyl butyric acid as well as 2μg/ml of tunicamycin respectively as negative and positive controls for inducing ER-stress. We measured the viability of treated cells by the MTT test. Besides, the expression levels of TLR4 gene and protein were evaluated using western blotting and reverse transcription- quantitative polymerase chain reaction (RT-qPCR) analysis. One-Way ANOVA was used for statistical analysis.

No cell death was observed in treated cells. The cells treated with 150 and 300 μM doses of TMAO for 24 hours showed a significant elevation in the protein and/or mRNA levels of TLR4 when compared to normal control or tunicamycin-treated cells.

Our results may in part elucidate the mechanism by which TMAO induces the macrophage inflammatory reactions in response to the induction of ER stress, similar to what happens during atherosclerosis. It also provides documentation to support the direct contribution of TLR4 in TMAO-induced inflammation.

Despite the positive role of rotifers in many hatcheries for feeding the early stages of aquatic larvae, the lower mineral content of zinc (Zn) is one of the disadvantages of rotifer compared to copepods. Therefore, it is necessary to increase its amounts through enrichment. For this purpose, in the present study, a combination of algae Isochrysis aff. galbana and Nannochloropsis oculata were enriched with zinc sulfate for 1 and 3 hours. Due to obtaining better results in 1 hour, its effects on the growth and enrichment of rotifer were surveyed. The 1: 1 alga composition was enriched with zinc sulfate at concentrations (45, 90, and 135 mg/l). The highest amount of zinc was observed in the mixed algal enriched with 90 mg/l for 1 h, which had the highest copper amount and there was no significant difference with the control group. Also, the manganese amount was higher than the other treatments except for the control group. After feeding the rotifers with enriched algae for 1 hour, the best treatment was 45 mg/l, which also contained the second level of zinc and the first level of manganese, copper, potassium, and sodium. On the other hand, this treatment had the highest number of eggs on the peak day of reproduction treatments (third day) and its population density in the last days was not significantly different from the control group (p <0.05). Zn-enriched rotifers can be used to feed marine fish to meet the nutritional needs of aquatic larvae.

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.

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).

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.

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. 

Despite the positive role of rotifers in many hatcheries for feeding the early stages of fish larvae and crustaceans, the lower content of minerals such as selenium (Se, 30 times) and zinc (Zn, 5 times) is considered as one of the disadvantages of this live prey compared to copepods. Therefore, increasing the amount of these nutrients through enrichment is essential. To this end, the present study investigated the effects of the combination of 2 algae Isochrysis aff. galbana and Nannochloropsis oculata enriched with sodium selenite and zinc sulfate on growth and mineral content of rotifer. First, the 1:1 composition of the algae was enriched with a mixture of zinc and selenium salts (each with concentrations of 20, 40 and 80 mg L-1 in algal enrichment medium). The highest number of rotifers (339 ± 11.06 ind mL-1), SGR (± 0.48 ± 0.008 day-1), and the lowest doubling time without any significant difference observed in rotifers fed with algae enriched with 40 mg L-1 of both minerals (p>0.05). However, the highest amount of Zn and Se were observed in those fed with 80 mg L-1 of both minerals. In conclusion, feeding rotifers with Se and Zn enriched algae increased Se and Zn in the rotifers. Thus, microalgae enriched with Se and Zn can be used to feed marine rotifer to meet the nutritional requirements.

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.

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.

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).

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.

Lifestyle modification is an important aspect of preventing various diseases, including various types of cancer. The aim of the present study was to investigate the effect of eight weeks of aerobic training and green tea extract on NF-κB, COX-2 and suppressor protein of p53 in prostate tissue of healthy rats.

Materials & Methods:

32 male Wistar rats were randomly divided into four groups: green tea extract, exercise training, green tea extract + exercise training, and control. The exercise training program included aerobic training on a low to moderate intensity on the treadmill (at a speed of 3-10 m/s, 3 sets of 15 min with 2 min of rest between sets, 5 days a week). Green tea extract was gavaged at a dose of 1.3 ml of solution at a concentration of 10 mg/100 ml (3 sessions per week). Scarify was performed 48 hours after the end of the intervention. The results were analyzed using parametric statistical methods of analysis of covariance and one way ANOVA.

Aerobic training significantly increased NF-κB level compared to the control group (P = 0.02). The combination of aerobic training and consumption of green tea extract did not significantly change the level of NF-κB. The COX-2 and p53 levels were not significantly different after aerobic training and green tea extract (P > 0.05).

Green tea supplementation seems to modulate NF-κB level following aerobic training, which can reflect the anti-inflammatory effects of green tea and its role in preventing prostate cancer.

There are promising data that exercise training and dietary polyphenols intake may alter the tumor microenvironment to facilitate conventional treatment.

The purpose of this study was to investigate the effect of eight weeks of aerobic training (AT) and green tea extract (GTE) consumption on matrix metalloproteinases-2 and -9 (MMP-2/-9) and vascular endothelial growth factor (VEGF) in normal and prostate cancer tissue of rats.

Ninety Wistar rats were assigned to two healthy and cancer groups, then the healthy group was divided into four subgroups, including healthy control (Ctr), healthy AT (treadmill exercise for 5 days/week), healthy GTE (three times of week for eight weeks by gavage) and healthy AT + GTE, and after tumor induction, the cancer group was divided into five subgroups, including cancer control (CaCtr), cancer AT (CaAT), cancer GTE (CaGTE), cancer AT + GTE and sham groups.

The results showed that there was no significant difference between healthy and CaCtr groups in the MMP-2 levels (P = 0.07), but MMP-2 levels in the CaAT group were lower than the healthy AT (P = 0.01). No significant changes were found in the levels of MMP-9 and VEGF between the healthy and cancer groups (P = 0.23 and P = 0.08, respectively).

The present study demonstrated that low to moderate-intensity aerobic training and treatment with a low dose of green tea extract did not change angiogenesis and metastasis markers. These results pave the way for further researches on modulation approaches for tumor metastasis and vascularization in responses to exercise training and antioxidant supplementation

Nonalcoholic fatty liver disease (NAFLD) and gallstone disease (GD) are both highly prevalent in the general population and have many risk factors in common. The aim of this study was to evaluate the prevalence of GD in the patients with NAFLD.

In this case-control study, our case group included 145 patients with NAFLD and control group consisted of 215 age-, sex-, and BMI-matched healthy subjects. NAFLD and gallstone disease were diagnosed by sonography. The prevalence and risk factors for gallstone disease were evaluated in the groups and compared between the 2 groups.

The frequency rates of gallstones in the case and control groups were 10/135 (6.9 %) and 5/210 (2.3 %) respectively (p=0.04). The frequency rates of gallstones in the female participants in the case and control groups were 6.8% and 3.4%, respectively, and in the men were 6.9% in the case and 1% in the control groups. We found no significant relationship between sex and frequency of gallstones in both groups (P> 0.05). In the control group, the prevalence rate of GD in the subjects over 50 years of age were significantly higher than that in the subjects under 50 years of age (P = 0.04). However, there was no significant difference between ag e and GD in the case group (P = 0.51). Also, in both groups, the prevalence of GD was significantly higher in obese subjects (BMI> 30) (P> 0.05). There was a significant difference between the grade of fatty liver and the prevalence of GD (P = 0.01). Comparison of the risk factors associated with the GD, showed no significant difference between the two groups.

The prevalence of gallstones was more in NAFLD than in normal population and was associated with grade of fatty liver disease.

Mesenchymal stem cells (MSCs) can be used to treat premature ovarian failure (POF). Different methods have already been applied to detect MSCs in tissues. This study aimed to investigate the quantitative distribution of CM-DiI-labeled human umbilical cord vein MSCs (hUCV-MSCs) in different regions of the ovarian tissue of the cyclophosphamide ( CTX )-induced POF in mice. Adult female C57BL/6 mice (n = 40) were divided into four groups: (1) Mice receiving PBS as control (Ctrl) group; (2) mice receiving hUCV-MSCs intravenously as Ctrl + hUCV-MSCs group; (3) mice receiving CTX intraperitoneally (i.p.) as CTX group; (4) mice receiving CM-DiI-labeled hUCV-MSCs after CTX injection as CTX + hUCV-MSCs group. Histological changes and CM-DiI-labeled hUCV-MSCs distribution were analyzed in the ovarian tissues. Quantitative real-time PCR was performed to detect human mitochondrial cytochrome b (MTCYB) gene in the ovarian tissues of the mice. The mean number of the fluorescent hUCV-MSCs was 20 ± 2.5 (57.1%) in the medulla, 11.3 ± 2.8 (32.2%) in the cortex, and 5.5 ± 1 (15%) in the germinal epithelium of the ovarian tissue (p < 0.05). Moreover, MTCYB gene was detected in the mice ovaries of the CTX + hUCV-MSCs group, but not in other groups. Our findings suggest that the distribution of the transplanted hUCV-MSCs in different regions of the ovarian tissue is not equal, and it is greater in the medulla than the cortex and germinal epithelium. This is the first report of quantitative distribution of MSCs in different regions of ovarian tissue in the POF model.