Expression levels of the BDNF gene and histone modifications around its promoters in the ventral tegmental area and locus ceruleus of rats during forced abstinence from morphine.

Brain-derived neurotrophic factor (BDNF) plays a role in mediating molecular, cellular, and behavioral adaptations underlying drug addiction. Here, we examined the influence of withdrawal from repeated morphine treatment on the expression of BDNF mRNA in the ventral tegmental area (VTA) and locus coeruleus (LC) of the rat brain. We also studied whether alternations in mRNA levels of BDNF in these tissues are associated with histone modifications around promoters II and III of the BDNF gene. Thus, chromatin immunoprecipitation (CHIP) and quantitative (q)-PCR were employed to assess acetylation of histone H3 at K9/K14 and trimethylation of histone H3 at K9. Results of qRT-PCR showed that levels of BDNF mRNA in both VTA and LC were significantly increased 7 days rather than 2 h or 24 h following the last injection of morphine. Consistently, CHIP and qPCR analysis revealed that on day 7 of morphine abstinence, both VTA and LC levels of histone methylation at BDNF promoters II and III of morphine treated rats were significantly lower than control animals. Morphine withdrawal caused only a significant increase in H3 acetylation at the promoter II in the LC. These data demonstrate the involvement of histone H3 methylation in the regulation of gene expression in the VTA and LC of rats during forced abstinence of morphine.

Anti-Proliferative and Anti-Telomerase Effects of Blackberry Juice and Berry- Derived Polyphenols on HepG2 Liver Cancer Cells and Normal Human Blood Mononuclear Cells.

BACKGROUND: Previous studies have provided strong evidence for the anticancer activity of berry fruits. OBJECTIVE: In this study, we investigated the effects of blackberry juice and three berry- polyphenolic compounds on cell proliferation and telomerase activity in human hepatoma HepG2 and normal peripheral blood mononuclear cells (PBMCs). METHODS: The cell viability and telomerase activity were measured by MTT and TRAP assay, respectively. Berry effects on the expression of genes were determined by quantitative RT-PCR assay. RESULTS: Blackberry, gallic acid, and resveratrol inhibited proliferation of both HepG2 and PBMC cells in a dosedependent manner. Resveratrol was more effective than gallic acid for reducing the viability of HepG2 cells, but both showed the same level of growth inhibition in PBMC cells. Berry, resveratrol, and gallic acid significantly inhibited telomerase activity in HepG2 cells. The antiproliferative effect of berry was associated with apoptotic DNA fragmentation. Gallic acid was more effective for reducing telomerase activity than resveratrol, but anthocyanin moderately increased telomerase activity in cancer cells. Telomerase activity was induced by all three polyphenols in PBMCs. Overall, Krumanin chloride was more effective to induce telomerase than gallic acid and resveratrol in PBMC cells. There was no significant difference in hTERT, hTR, and Dnmts expressions between berry treated and the control untreated HepG2 cells. But, a significant downregulation of HDAC1 and HDAC2 and upregulation of SIRT1 were observed in berry-treated cells. CONCLUSION: These data indicate that the berry anticancer effect is associated with antitelomerase activity and changes in HDACs expression. The data also suggest that berry antitelomerase activity is mainly related to its gallic acid and resveratrol, but not anthocyanin content.

Improving the endothelial dysfunction in type 2 diabetes with chromium and vitamin D3 byreducing homocysteine and oxidative stress: A randomized placebo-controlled trial.

BACKGROUND: Chromium picolinate (CrPic) and vitamin D3 are known as two antioxidant micronutrients. Through inducing endothelial dysfunction, oxidants such as homocysteine (Hct) and malondialdehyde (MDA) lead to cardiovascular disease in type 2 diabetes mellitus (T2DM). No published data has directly examined the effects of these two antioxidants on improving the endothelial dysfunction in T2DM throughreducing homocysteine and oxidative stress. METHODS: Subjects (n = 92) in this randomized, double blind, placebo-control study were randomly assigned to receive oral placebo (group I), D3 (group II: 50,000 IU/ week), chromium picolinate (CrPic) (group III: 500 µg/day), and both vitamin D3 and CrPic (group IV) for four months. Fasting blood samples were drawn at study baseline and following intervention to determine Hct, MDA, total antioxidant capacity (TAC), total thiol groups (SHs), vascular cell adhesion molecule- 1 (VCAM-1), and plasminogen activator inhibitor-1 (PAI-1). RESULTS: After intervention, MDA significantly decreased in groups II and IV; TAC significantly increased in group IV, and SHs significantly augmented in group III; Hct was significantly reduced in groups II, III, and IV; and VCAM-1 significantly decreased in groups III and IV and PAI-1 was significantly reduced in groups II, III, and IV. CONCLUSION: Our findings suggest that through reducing homocysteine and oxidative stress and improving endothelial dysfunction, chromium and vitamin D3 co-supplementation might be predictive and preventive of cardiovascular diseasesassociated with T2DM. IRCT, IRCT20190610043852N1, registered 21 October 2019, https://fa.irct.ir/user/trial/42293/view.

The effect of hydro-alcoholic extract of Ceratonia Silique L. on spermatogenesis index in rats treated with cyclophosphamide: An experimental study.

BACKGROUND: The aim of this study was to investigate the antioxidant effects of Ceratonia extract on improving the toxicity induced by cyclophosphamide (CP) on spermatogenesis. Materials and Methods: 54 male Wistar rats (4 months old) weighing 200-250 gr were randomly divided into 6 groups (n = 9/each). OBJECTIVE: "group 1 (control) underwent the normal diet and water; group 2 (sham) received 2 ml/day normal saline; group 3 (positive control) received 300 mg/kg/day Ceratonia extract; group 4 (Ceratonia + CP) received Ceratonia extract (300 mg/kg/day) + 5 mg/kg/day CP (Endoxan, baxter oncology gmbh, Germany) after 4 hr; group 5 (CP) received 5 mg/kg/day CP + normal saline 4 hr after it; and group 6 (CP + Ceratonia) received Ceratonia extract (300 mg/kg/day) 4 hr after 5 mg/kg/day CP." 24 hr after the last gavage, heart blood sampling was performed to measure the levels of malondialdehyde (MDA), ferric reducing antioxidant power, testosterone, luteinizing hormone, and follicle-stimulating hormone. The left caudal epididymis was cut in the Ham's F10 and the released spermatozoa were used to analyze sperm parameters. The histology of the right testes was studied using stereological techniques and the left testes were used to measure the level of tissue MDA and ferric reducing antioxidant power. RESULTS: A significant increase in the mean level of MDA (p = 0.013) was seen in the CP compared to the control group. Sperm motility (p = 0.001) and count (p = 0.002), serum and tissue total antioxidant (p ≤ 0.001) and serum testosterone levels (p = 0.019) decreased in the CP compared to the control group. Ceratonia extract could significantly prevent the adverse effects of CP on sperm motility (p < 0.001), the mean levels of tissue MDA (p = 0.018), serum total antioxidant (p = 0.045), and testosterone (p < 0.001). CONCLUSION: The Ceratonia extract can modify the reproductive toxicity of CP in rat due to the presence of antioxidant compounds.

Epithelial-to-mesenchymal transition in thyroid cancer: a comprehensive review.

The Metastatic progression of solid tumors, such as thyroid cancer is a complex process which involves various factors. Current understanding on the role of epithelial-mesenchymal transition (EMT) in thyroid carcinomas suggests that EMT is implicated in the progression from follicular thyroid cancer (FTC) and papillary thyroid cancer (PTC) to poorly differentiated thyroid carcinoma (PDTC) and anaplastic thyroid cancer (ATC). According to the literature, the initiation of the EMT program in thyroid epithelial cells elevates the number of stem cells, which contribute to recurrent and metastatic diseases. The EMT process is orchestrated by a complex network of transcription factors, growth factors, signaling cascades, epigenetic modulations, and the tumor milieu. These factors have been shown to be dysregulated in thyroid carcinomas. Therefore, molecular interferences restoring the expression of tumor suppressors, or thwarting overexpressed oncogenes is a hopeful therapeutic method to improve the treatment of progressive diseases. In this review, we summarize the recent findings on EMT in thyroid cancer focusing on the main role-players and regulators of this process in thyroid tumors.

Acriflavine enhances the antitumor activity of the chemotherapeutic drug 5-fluorouracil in colorectal cancer cells.

5-Fluorouracil (5-FU)-based chemotherapy improves the overall survival rates of patients with colorectal cancer (CRC). However, only a small proportion of patients respond to 5-FU when used as a single agent. The aim of the present study was to investigate whether the anticancer property of 5-FU is potentiated by combination treatment with acriflavine (ACF) in CRC cells. Additionally, the potential underlying molecular mechanisms of the cytotoxic effect of ACF were determined. The cytotoxic effects of ACF, 5-FU and irinotecan on different CRC cell lines with different p53 status were investigated using an MTT assay. SW480 cells that express a mutated form of p53 and two other CRC cell lines were used, HCT116 and LS174T, with wild-type p53. To determine the effect of ACF on the sensitivity of cells to 5-FU, cells were co-treated with the 30% maximal inhibitory concentration (IC30) of ACF and various concentrations of 5-FU, or pretreated with the IC30 of ACF and various concentrations of 5-FU. To assess the mechanism of action of ACF, cells were treated with IC30 values of the compound and then the reverse transcription-quantitative polymerase chain reaction was used to evaluate mRNA levels of hypoxia-inducible factor-1α (HIF-1α) and topoisomerase 2. Results indicate that pretreatment with ACF markedly sensitized CRC cells to the cytotoxic effects of 5-FU, whereas simultaneous treatment with ACF and 5-FU were not able to alter the resistance of CRC cells to 5-FU. In comparison with irinotecan, ACF was a more potent agent for enhancing the antitumor activity of 5-FU. ACF did not alter the mRNA levels of either HIF-1α or topoisomerase 2. The results of the present study reveal for the first time that pretreatment of CRC cells with ACF markedly increases the cytotoxic effects of 5-FU, regardless of the p53 status of cells.

Resveratrol decreases apoptosis and NLRP3 complex expressions in experimental varicocele rat model.

OBJECTIVES: Varicocele is an abnormal dilation in the testicular vein, which can cause hypoxia, reactive oxygen species accumulation, elevation in testicular temperature, and promote apoptosis and increase proinflammatory cytokine production. According to the varicocele pathophysiology, it is possible that a group of cytosolic receptors called nucleotide oligomerization domain (NOD)-like receptor family pyrin domain containing 3 (NLRP3) inflammasomes also involve in varicocele pathogenesis. Due to the important role of antioxidant in decreasing the testis tissue damage, in this study we investigated the protective effect of resveratrol (RES) on NLRP3 complex and apoptosis in experimental varicocele rats. MATERIALS AND METHODS: In this study, 40 male Wistar rats were randomly divided into 5 groups (8 rats in each group): Control, experimental left varicocele (ELV), ELV + ethanol, ELV + 20 mg/kg RES and ELV + 50 mg/kg RES. Varicocele was induced by partial ligation of the left renal vein. Three months after varicocele induction, RESwas orally administered to rats for 1 month. The expression levels of NLRP3, apoptosis associated speck-like protein (ASC), caspase-1, Bax and Bcl2 were analyzed using real time PCR. RESULTS: Our results showed that RESat both doses significantly (P≤ 0.05) decreased the gene expression levels of ASC, NLRP3, caspase-1 and Bax and increased Bcl2 gene expression at high dose. CONCLUSIONS: RESby reducing inflammatory factors and decreasing apoptosis might be used as adjuvant therapy to reduce varicocele complication.

Association between chemerin rs17173608 and rs4721 gene polymorphisms and gestational diabetes mellitus in Iranian pregnant women.

Gestational diabetes mellitus (GDM) is defined as hyperglycemia detected during pregnancy and its risk is increased with obesity. Chemerin, an adipokine, has been proposed as potential mediators of insulin resistance in GDM. This case-control study was designed to assess the relation between chemerin SNPs rs4721 (or rs10278590) and rs17173608 and the development of GDM. One hundred thirty GDM pregnant women with GDM and 160 healthy pregnant women were enrolled in this study. The diagnosis of GDM was based on the International Association of Diabetes and Pregnancy Study Group (IADPSG) criteria. Chemerin rs4721 polymorphism gene was amplified through PCR, and SNP was detected using restriction enzyme AluI. Genotyping for chemerin rs17173608 polymorphism was performed by using tetra-amplification refractory mutation system polymerase chain reaction (T-ARMS-PCR). Blood glucose level was measured by an enzymatic method. Our finding showed that the genotypes frequency of chemerin rs4721 polymorphism was significantly different between GDM and non-GDM groups (χ2 = 7.44, P = 0.02). The genotype of rs4721 was significantly associated with GDM in co-dominant and dominant genotypes (GG vs GT, OR = 2.3, 95%CI = 1.24-4.24, P = 0.008, and GG vs GT + TT, OR = 2.21, 95%CI = 1.23-3.99, P = 0.008, respectively). No significant difference was observed in allele frequency between case and control groups (P = 0.62). Moreover, the genotypes and allele frequencies of chemerin rs17173608 polymorphism did not show significant differences between GDM and non-GDM (P > 0.05). We concluded that the genotype of rs4721 was found to contribute significant risk to GDM while genotype of rs17173608 could not predict the risk of GDM.

Effects of different Sertoli cell types on the maintenance of adult spermatogonial stem cells in vitro.

Spermatongonial stem cells (SSCs) are unique testis cells that are able to proliferate, differentiate, and transmit genetic information to the next generation. However, the effect of different Sertoli cell types on the expression of specific SSC genes is not yet well understood. In this study, we compare the in vitro effect of adult Sertoli cells, embryonic Sertoli cells, and TM4 (a Sertoli cell line) as feeder layers on the expression of SSC genes. SSCs were isolated from the testis of adult male mice and purified by differential plating. Following enrichment, SSCs were cultivated for 1 and 2 wk in the presence of various feeders. The expression of SSC-specific genes (Mvh, ZBTB, and c-kit) was evaluated by real-time polymerase chain reaction. Our results revealed that expression of the specific SSC genes was significantly higher in the embryonic Sertoli cells after 1 and 2 wk compared to the adult Sertoli cells and the TM4 group. Our finding suggest that co-culturing of SSCs with embryonic Sertoli cells is helpful for in vitro cultivation of SSCs and might improve the self-renewal of these stem cells.

Association between vitamin D receptor ApaI and TaqI gene polymorphisms and gestational diabetes mellitus in an Iranian pregnant women population.

It has been proposed that variants of the nuclear vitamin D receptor (VDR) gene are associated with a susceptibility to type 2 diabetes mellitus (T2DM) and gestational diabetes mellitus (GDM). Our study was aimed to evaluate a possible association between the VDR ApaI (rs7975232) and TaqI (rs731236) gene polymorphisms and susceptibility to GDM in an Iranian pregnant women population. This case-control study was performed on a population of pregnant Iranian women, including 157 GDM and 157 non-GDM subjects.VDR ApaI and TaqI polymorphisms were assessed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).Our finding showed that the genotypes frequency of VDR ApaI polymorphism was significantly different between GDM and non-GDM groups (χ(2)=8.5, P=0.014). The CC genotype increased the risk of GDM as compared to the AA genotype (AA vs.CC, OR=2.996, 95% CI=1.278-7.022, P=0.012). The genotype and allele frequencies of VDR TaqI polymorphism were significantly different between GDM and non-GDM subjects (χ(2)=7.27, P=0.026, χ(2)=4.08, P=0.043 respectively). A significant protection was shown against GDM in VDR TaqI genotypes and allele (TT vs.TC, OR=0.523, 95% CI=0.23-0.84, P=0.007, TT vs. TC+CC, OR=0.546, 95% CI=0.35-0.86, P=0.009, T vs. C, OR=0.711, 95% CI=0.511-0.99, P=0.043). In conclusion, our findings show a significant association between VDR ApaI and TaqI gene polymorphisms and the GDM at the investigated loci.

Oxidative stress in acute human poisoning with organophosphorus insecticides; a case control study.

Free radicals play an important role in toxicity of pesticides and environmental chemicals. Organophosphorus insecticides (OPIs) may induce oxidative stress leading to generation of free radicals and alteration in antioxidant system. To complete the previous surveys, this study was conducted to evaluate the existence of oxidative stress, balance between total antioxidant capacity and oxygen free radicals in patients with acute OPI exposure. In this case control study, a total of 22 acute OPI poisoning patients were included and blood samples were analyzed for lipid peroxidation, total antioxidant capacity, total thiol groups, and cholinesterase levels. The results showed significant lipid peroxidation accompanied with decreased levels of total antioxidant capacity, total thiols, and cholinesterase activity. A significant correlation existed between cholinesterase depression and reduced total antioxidant capacity. It is concluded that oxygen free radicals and their related interactions like lipid peroxidation are present in acute OPI poisoning. Use of antioxidants may be beneficial in treatment of OPIs acute poisoning which remains to be elucidated by further clinical trials.