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Introduction: MicroRNAs (miRs) are a group of endogenous, non-coding, 18-24 nucleotide length single-strand RNAs that mediate gene expression at the post-transcriptional level through mRNA degradation or translational repression. They are involved in regulating diverse cellular biological processes such as cell cycle, differentiation, and apoptosis. The deregulation of miRs affects normal biological processes, leading to malignancies, including oral squamous cell carcinoma (OSCC). This study evaluates the expression level of miR-21-5p and miR-429 genes in biopsy samples from patients with OSCC and performs a comparison with controls. Materials and Methods: In this study, tissue samples were obtained from 40 individuals (20 OSCC patients and 20 healthy controls) to determine miR-21-5p and miR-429 expression using the ΔCT method and analyzed by the Mann–Whitney test. Results: The mean age of subjects in the control and patient groups was 47.15 and 53.8 years, respectively. According to the Mann–Whitney test, significant differences were observed in miR-21-5p (p < 0.0001) and miR-429 (p = 0.0191) expression levels between the two groups (p < 0.05). Conclusions: The expression of miR-21-5p, miR-429, and combined miRNAs in the OSCC group was significantly higher compared to the control group. As a result, changes in the expression of these biomarkers in cancerous tissues could potentially be considered as a marker for the early diagnosis of OSCC. [ABSTRACT FROM AUTHOR]

Hemojuvelin (HJV) enhances signaling to the iron hormone hepcidin and its deficiency causes iron overload, a risk factor for hepatocellular carcinoma (HCC). We utilized Hjv−/− mice to dissect mechanisms for hepatocarcinogenesis. We show that suboptimal treatment with diethylnitrosamine (DEN) triggers HCC only in Hjv−/− but not wt mice. Liver proteomics data were obtained by mass spectrometry. Hierarchical clustering analysis revealed that Hjv deficiency and DEN elicit similar liver proteomic responses, including induction of mitochondrial proteins. Dietary iron overload of wt mice does not recapitulate the liver proteomic phenotype of Hjv−/− animals, which is only partially corrected by iron depletion. Consistent with these data, primary Hjv−/− hepatocytes exhibit mitochondrial hyperactivity, while aged Hjv−/− mice develop spontaneous HCC. Moreover, low expression of HJV or hepcidin (HAMP) mRNAs predicts poor prognosis in HCC patients. We conclude that Hjv has a hepatoprotective function and its deficiency in mice promotes mitochondrial dysfunction and hepatocarcinogenesis. Hemojuvelin (HJV), a BMP co-receptor promoting hepcidin expression in the liver, has a hepatoprotective function and its deficiency in mice triggers mitochondrial dysfunction and hepatocarcinogenesis. [ABSTRACT FROM AUTHOR]

Concurrent exposure to styrene (ST) and noise is common especially in industrial environments. The present study aims to determine the related oxidant-induced changes as the result of combined exposure to ST and noise. For this purpose, 24 male Wistar rats were used in four experimental groups (n = 6/groups): (1) control group, (2) the group exposed to an octave band of noise centered at 8 kHz (100 dB SPL) (6 h/day), (3) the group inhalationally exposed to ST (750 ppm) (6 h/day), (4) the group exposed to noise and ST simultaneously. The DNA damage was measured by assessing the concentration of 8-hydroxyl-2-deoxyguanosine (8-OHdG) using ELISA kit. Levels of lipid peroxidation (MDA), GSH and antioxidative activity of SOD and CAT were also determined in whole lung tissues. The results relatively indicated that sub-acute exposure to both noise and ST can lead to pathological damage in rat lung tissues. Furthermore, enhanced levels of 8-OHdG and MDA production were observed in lung tissues. In contrast, GSH, CAT and SOD were markedly reduced in co-exposed group. The results of the study verified additive interaction between noise and ST on accumulation of DNA oxidation products, progressive morphological damages as well as undermining the antioxidative defense system in the rat lung tissues. [ABSTRACT FROM AUTHOR]

The aim of this study was to examine the impact of intake of a lactic acid bacterium, Lactobacillus plantarum 299v (Lp299v) on aflatoxin-induced hepatotoxicity in broilers. For this, broilers were intoxicated with dietary aflatoxins and simultaneously treated with live Lp299v in drinking water. One-day-old male broilers were divided into eight groups (n = 10/group) as follows. Aflatoxin groups fed basal diet contaminated with aflatoxins (200 or 2000 ppb). The probiotic groups received drinking water enriched with live (Lp299v) (108 CFU/ml). A group of birds was given a commercial mycotoxin binder (2.5 g/kg feed). Control groups received basal diet without probiotic or aflatoxin binder. The growth performance was calculated for the entire period (0–42 days), and blood and liver specimens were processed for histology and determination of liver damage markers. Results showed extensive damage including bile duct hyperplasia, hepatocellular ballooning, and necrosis in chickens fed aflatoxin alone. However, liver lesions were limited to lobular inflammation and pyknosis in broilers treated with aflatoxins along with Lp299v. The histology of the liver tissues from the birds on aflatoxin-free diet + probiotic appeared to be normal when compared to the respective controls. Histopathological indices in different experimental groups were corroborated with the liver damage markers namely, serum ALT, AST, LD, and γ-GT. It is concluded that the improvement in the growth performance and prevention of aflatoxin-related liver lesions could be mainly assigned to the probiotic therapy for the entire period of breeding, although the aflatoxin-binding ability of the Lp299v in inactivation of aflatoxins cannot be ruled out. RESEARCH HIGHLIGHTS Aflatoxin-related liver damage progression was inhibited by probiotics in broilers. Aflatoxin inactivation by probiotics can be assessed by liver histopathology. Growth performance in broilers was improved following the intake of probiotics. [ABSTRACT FROM AUTHOR]

Aim: Due to lipases' regio-selectivity and ability to catalyze different reactions such as hydrolysis, esterification, and transesterification, the enzyme is attractive in biotransformation technology. Besides, another technology, namely enzyme immobilization, has attracted scientists/technologists' attention to employ immobilized lipase in such a field. Thus lipase of Candida rugosa was immobilized onto silica nanoparticles through adsorption. Furthermore, the immobilized biocatalyst was characterized and used to esterify ibuprofen enantioselectively. Methods: To characterize immobilized lipase onto silica nanoparticles scanning electron microscopy (SEM) and dynamic light scattering (DLS) were used. Results: The catalytic properties of both immobilized and free lipases such as optima pH and temperature were not different. According to the results, the immobilized lipase on silica nanoparticles showed 45% and 96% conversion (C) and enantioselectivity (ees), respectively. In comparison to free lipase, the immobilized enzyme came with better catalytic activity. Conclusion: Silica nanoparticles as one of the most promising materials for the immobilization of lipase in enantioselective esterification of ibuprofen, were introduced in this work. [ABSTRACT FROM AUTHOR]

Objective(s): Autophagy is an intracellular degradation system of damaged proteins and organelles; however, the role of autophagy in the progression of cancer remains unclear. In recent years, mesenchymal stem cell (MSC)-based approaches have attracted considerable attention for anti-cancer therapy. The present study aimed to examine the interaction of MSCs with the breast cancer cells under autophagy-induced conditions. Materials and Methods: In this study, MSCs isolated from human adipose tissue were co-cultured with MDA-MB 231, a breast cancer cell line, and the autophagy process was induced by tunicamycin treatment. The cell viability was monitored by the MTT assay, and the cells were recovered at different time intervals (24 or 48 hours) to determine autophagy markers such as Beclin, mTOR and the ratio of LC3II/I expression. Additionally, the animal study was conducted using a mouse model of breast cancer treated with isogenic adipose-derived MSCs, and the expression of Beclin and Ki67 was determined using immunohistochemistry in breast tumor tissue. Results: In cancer cells co-cultured with MSCs, the cell proliferation was increased, the Beclin expression and the LC3II/I protein ratio were decreased, and the mTOR expression was increased in MDA-MB 231 upon co-cultured with MSCs. Direct injection of MSCs to a mouse model of breast cancer showed an increase in tumor volume, an increase in the accumulation of Ki67 and a decrease in the Beclin expression in tumor tissues. Conclusion: The data may suggest that suppressed autophagy in breast cancer cells is probably a mechanism by which MSCs can induce cancer cell proliferation. [ABSTRACT FROM AUTHOR]

This study was carried out to examine the role of intracellular albumin in the modulation of oxidative damage induced by glutathione modifiers in HepG2 cells. Also, the relationship of albumin synthesis with oxidative stress factors including antioxidants was studied. HepG2 cell culture was supplemented with glutathione modifiers; L‐Buthionine‐sulfoximine (BSO; 0.1 and 1.0 mM) or N‐acetyl cysteine (NAC; 1 and 10 mM) and the cell viability and changes in reduced glutathione (GSH), oxidized glutathione (GSSG), reactive oxygen species (ROS), catalase, and superoxide dismutase were measured. Besides, albumin expression at protein and mRNA levels was determined in cells pretreated with BSO or NAC. Kinetic studies showed that albumin expression in HepG2 cells is correlated with GSH and GSSG levels. Changes in albumin expression at protein and mRNA levels reached their maximum (19% and 55%, respectively) in the cells 6 H after NAC treatments. A substantial decrease in intracellular albumin due to BSO (27%) was associated with a significant increase in the generation of cellular ROS (17%). In contrast, increased albumin synthesis (intracellular and secretory) was associated with inhibition in cellular ROS. Overall data may suggest that albumin expression in coordination with the glutathione redox system is part of the antioxidant defense mechanism in liver cells. [ABSTRACT FROM AUTHOR]

In recent years, a lot of attention has been paid to quantum dot (QD) nanoparticles as fluorescent sensors for sensitive and accurate detection of cancer biomarkers. Here, using a homemade specific monoclonal antibody against CA125 and QD525- or FITC-labeled probes, expression of this marker in an ovarian cancer cell line and cancer tissues were traced and optical properties of fluorophores were compared qualitatively and quantitatively. Our results clearly showed that besides lower background and exceptionally higher photobleaching resistance, QD525 exhibited higher fluorescent intensity for both ovarian cancer cell and tissues at different exposure times (p<0.0001) and excitation filter sets (p<0.0001) exemplified by significantly higher staining index (p<0.016). More importantly, the FITC-labeled probe detected antigen-antibody complex at minimum concentration of 0.3 μg/mL of anti-CA125, while reactivity limit decreased to 0.078 &3956;g/mL of anti-CA125 when QD525-labeled probe was applied showing four times higher reactivity level of QD525 probe compared to the same probe labeled with FITC. Based on our results, it seems that QDs are inimitable tags for sensitive detection and localization of ovarian cancer micrometastasis and molecular demarcation of cancer tissues in surgical practice, which subsequently figure out accurate therapeutic approaches. [ABSTRACT FROM AUTHOR]

Resistance to oxidative damages in undifferentiated mesenchymal stem cells (MSCs) in comparison with the undifferentiated progenitor cells may differ depending on several factors. This study was carried out to examine the impact of hepatogenic differentiation process of MSCs on oxidative DNA damage markers. Hepatic differentiation of MSCs was carried out using a two-step conventional protocol and the cells were processed for characterization using morphological and biochemical markers. During the course of differentiation cellular levels of reactive oxygen species (ROS), 8-hydroxy-2'-deoxyguanosine (8-OH-dG) and expression of ataxia-telangiectasia mutated (ATM) protein were estimated at time intervals (10, 20 and 30 days). The results showed a decrease in cellular ROS (13%, P < 0.05) at early stages of hepatogenic differentiation. Similarly, there was a small but significant decrease in 8-OH-dG level and ATM expression in differentiated hepatocytes. Despite the small changes in oxidative damage factors and ATM expression during the differentiation process, the hepatocytes obtained were morphologically and biologically intact. [ABSTRACT FROM AUTHOR]

Objective: In vivo and in vitro stem cell differentiation into endothelial cells is a promising area of research for tissue engineering and cell therapy. Materials and Methods: We induced human mesenchymal stem cells (MSCs) to differentiate to endothelial cells that had the ability to form capillaries on an extracellular matrix (ECM) gel. Thereafter, the differentiated endothelial cells at early stage were characterized by expression of specific markers such as von Willebrand factor (vWF), vascular endothelial growth factor (VEGF) receptor 2, and CD31. In this experimental model, the endothelial cells were transplanted into the groins of severe combined immunodeficiency (SCID) mice. After 30 days, we obtained tissue biopsies from the transplantation sites. Biopsies were processed for histopathological and double immunohistochemistry (DIHC) staining. Results: Endothelial cells at the early stage of differentiation expressed endothelial markers. Hematoxylin and eosin (H&E) staining, in addition to DIHC demonstrated homing of the endothelial cells that underwent vascularization in the injected site. Conclusion: The data clearly showed that endothelial cells at the early stage of differentiation underwent neovascularization in vivo in SCID mice. Endothelial cells at their early stage of differentiation have been proven to be efficient for treatment of diseases with impaired vasculogenesis. [ABSTRACT FROM AUTHOR]

Apolipoprotein E (APOE) with its extraordinary features is readily assembled with hydrophobic compounds via its compact hydrophobic units (CHUs). These assemblies can then be converted to stable particles by protein–protein interactions via coiled coil regions (CCRs) which exist in APOE structure. Applying these features of APOE, we prepared novel nanoparticles called NAPOE, using no cross-linker. Vitamin D3 – a hydrophobic antitumor model – was loaded within the nanoparticles (NPs). The NPs were mostly spherical with the mean diameter and zeta potential of 94.39 ± 5.71 nm and −20 ± 0.3 mV, respectively. The molar ratio of VD3/APOE in NPs was 37.2 ± 0.61. The NPs targeted C6 glioma cellsin vitrovia over-expressed LDLRs. The efficiency of the NPs uptake to malignant C6 glioma cells was remarkable compared to non-tumor glial cells (p < 0.05). The releasing rate of hydrophobic cargo from the particles was high (p < 0.05) and reached to maximum, 12 h after targeting C6 cells. The size and drug loading of NPs were found to be controlled by the definite numbers of CCRs and CHUs in APOE. In conclusion, it is suggested that NAPOE NPs can facilitate the controlled delivery of hydrophobic drugs to the malignant C6 glioma cells according to the degree of invasiveness. [ABSTRACT FROM PUBLISHER]

Recent studies have been established high degree of genetic diversity in solid organ tumors among individuals and even between individual tumor cells. This intratumor and intertumor genetic diversity results in a heterogeneous tumor with unique characteristics which potentially allows effective drug therapy. The goal of pharmacogenomics is to elucidate the genetic network(s) that underlie drug efficacy and drug resistance. Advances in targeted and personalized therapy play an increasingly important role in many common cancers, notably lung cancer, due to the high incidence, prevalence, mortality and the greater tendency towards drug resistance seen in these patients. Non-small cell lung cancer (NSCLC) is characterized by mutations in the epidermal growth factor receptor (EGFR) and or downstream kinase pathways. This has led to the development of highly selective monoclonal antibodies and EGFR tyrosine kinase inhibitors (EGFR-TKIs) to prevent cancer initiation, proliferation, differentiation, angiogenesis, survival, and invasion. However, resistance to many of these new treatments is induced and further pharmacogenomic analysis has revealed mutations associated with increased or reduced drug efficacy. Combinations of kinase inhibitors or potentially the targeting of cancer stem cells may further increase the success of pharmacogenomics in treating patients with lung cancer. [ABSTRACT FROM AUTHOR]

Differentiation of mesenchymal stem cells (MSCs) to hepatocyte-like cells is associated with morphological and biological changes. In this study, the effect of hepatogenic differentiation on fatty acid profile and the expression of proliferator-activated receptors- α (PPAR- α) have been studied. For this purpose, MSCs isolated from human umbilical cord were differentiated into hepatocyte-like cells on selective culture media. The morphological and biochemical changes, PPAR- α expression and reactive oxygen species (ROS) levels were studied during the differentiation process. Besides, the cells were processed to determine changes in fatty acid profile using gas chromatography analysis. The results showed that hepatic differentiation of the MSCs is associated with a decrease in major polyunsaturated fatty acids in mature hepatocytes, whereas there was an increase in the saturated fatty acid (SFA) levels during hepatocyte maturation. The differentiation-dependent shift in the ratio of SFA/USFA was associated with changes in albumin and PPAR- α expression, whereas changes in fatty acid profile were independent of ROS production and lipid peroxidation in differentiating cells. In conclusion, these data may suggest that hepatocyte formation during the stem cell differentiation is associated with a shift in the fatty acid profile that is probably a normal phenomenon in hepatogenic differentiation of the MSCs. Copyright © 2014 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Differentiation of mesenchymal stem cells (MSCs) to hepatocytes-like cells is associated with alteration in the level of reactive oxygen species (ROS) and antioxidant defense system. Here, we report the role of glutathione in the functions of hepatocytes derived from MSCs. The stem cells undergoing differentiation were treated with glutathione modifiers [buthionine sulfoxide (BSO) or N-acetyl cysteine (NAC)], and hepatocytes were collected on day 14 of differentiation and analysed for their biological and metabolic functions. Differentiation process has been performed in presence of glutathione modifiers viz. BSO and NAC. Depending on the level of cellular glutathione, the proliferation rate of MSCs was affected. Glutathione depletion by BSO resulted in increased levels of albumin and ROS in hepatocytes. Whereas, albumin and ROS were inhibited in cells treated with glutathione precursor (NAC). The metabolic function of hepatocytes was elevated in BSO-treated cells as judged by increased urea, transferrin, albumin, alanine transaminase and aspartate transaminase secretions in the media. However, the metabolic activity of the hepatocytes was inhibited when glutathione was increased by NAC. We conclude that the efficiency of metabolic function of hepatocytes is inversely related to the levels of cellular glutathione. These data may suggest a novel role of glutathione in regulation of metabolic function of hepatocytes. Copyright © 2013 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Differentiation of human mesenchymal stem cells (MSCs) to metabolically active hepatocytes depends on different regulatory factors. Trans-differentiation of stem cells into specific cell lineage in the presence of specific stimuli is associated with the molecular and cellular damage. The aim of the present study was to examine the role of P53 in the regulation of cyclooxygenase-2 (COX-2) expression and the generation of protein and lipid oxidation during trans-differentiation of MSCs into hepatocyte-like cells. During the 3-week differentiation process of MSCs to hepatocyte-like cells we found that expression liver-specific markers was associated with increased levels of lipid peroxidation and protein carbonyl formation. Expression of P53 and COX-2 at mRNA and protein levels were evaluated in MSCs before and after differentiation on days 7, 14 and 21. We showed that the up-regulation of COX-2 was associated with augmentation of the rate of cell proliferation, morphological and biochemical changes of hepatocytes-like cells. However, in parallel the P53 at the mRNA level was down-regulated, and at protein levels accumulation in the nuclei was reduced during the hepatogenic differentiation time. Our results may suggest a P53-COX-2 pathway in the regulation of hepatogenic differentiation of stem cells, which is linked to differentiation-dependent molecular oxidative damage. [ABSTRACT FROM AUTHOR]

Objective: Multiple sclerosis (MS) is a chronic autoimmune disease due to demyelination of the central nervous system. It is believed that cytokines are involved in the pathogenesis of MS. The interleukin-2 (IL2) gene is powerful functional candidate that is involved in immune regulation and operation. In this study, for the first time, we investigated the effect of -475 A/T and -631 G/A IL2 polymorphisms on MS disease in Iranian patients. Materials and Methods: In this case-control study, 100 MS patients (mean age: 32.95 ± 6.51 years, age range: 20-42 years) selected according to McDonald criteria, and 100 ethnically, sex and age matched healthy controls (mean age: 29 ± 7.8 years, age range: 20-52 years) with no personal or family history of autoimmune diseases were studied. The restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) method was applied to define different alleles and genotypes of IL2 promoter single nucleotide polymorphism -475 A/T as well as -631 G/A among individuals. ?2 was calculated and Fisher's exact test was applied to analyze the obtained data. The value of p <0.05 was considered significantly . Results: Evaluation of the -475 IL2 revealed that T allele and A/T genotype are present in 2% and 4% of MS patients, respectively, whereas T allele was absent in control samples. The comparison between alleles and genotypes in MS patients and healthy controls was not significant (p=0.1). For the -631 position, 1% and 2% of MS patients carried A allele and A/G heterozygote genotypes, respectively. All control samples had G allele and G/G genotype. The differences between patients and controls were not significant (p=0.4). Moreover, our results showed a very low frequency of T at -475 and A at -631 IL2 position in each of the two groups. Conclusion: Both -475 and -631 IL2 polymorphisms were higher in MS patients as compared to controls, but the frequency differences were not significant. Based on these data, it is suggested that the -475 and -631 IL2 polymorphisms as functional promoter position may be involved in IL2 expression and regulation. To find out the exact effect of the mentioned SNPs on susceptibility to MS, study on a larger sample size is suggested. [ABSTRACT FROM AUTHOR]

Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system. Interleukin-2 (IL-2) is identified as the crucial and main immunoregulatory cytokines. Previously, we showed significant association between -330 T/T IL-2 genotype and relapsing remitting MS among Iranian population. In this study we investigated 100 relapsing remitting, 30 secondary progressive MS and 125 healthy controls to compare the relapsing remitting and secondary progressive course MS in association to -330 IL-2 polymorphism. Our results showed that the -330 T/T IL-2 genotype was significantly more frequent in relapsing remitting and secondary progressive MS than controls. The significant increased frequency of -330 T/T IL-2 genotype in secondary progressive than relapsing remitting MS, imply -330 T/T IL-2 genotype can cause higher susceptibility to secondary progressive MS than relapsing remitting. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to investigate the contribution of acute alcohol in sepsis-related liver damages using a Cecal Ligation and Puncture (CLP) model. Rats were divided into 7 groups (5 rats/group): control (saline-injected), sham-operated, CLP, ethanol (1.0 and 2.0g/kg b.w) and CLP+ethanol. The CLP+ethanol group received a single dose of ethanol following sepsis induction. Sepsis induction caused early changes in lipid peroxidation products in liver, whereas ethanol alone (2.0g/kg b.w) resulted in a significant increase (∼21%) in lipid peroxidation, which was further increased (∼57%) in CLP rats treated with alcohol. CLP operation and alcohol treatment exhibited additive effects on plasma catalase, liver glutathione and glutathione S-transferase (GST), which were primarily suppressed due to ethanol. Hepatic cytochrome P4501A1, which was elevated in CLP rats, was reversed in the CLP+ethanol group. Plasma tumor necrosis factor-α was markedly elevated (∼85%) in septic rats, but was unaffected in septic rats having received ethanol. Histopathological observations revealed that inflammatory reactions in liver in response to CLP operation are not intensified by ethanol administration. On the basis of biochemical and histopathological results, it can be concluded that acute ethanol treatment is responsible for early changes in oxidative stress, which may lead to polymicrobial sepsis-related organ damage. [Copyright &y& Elsevier]

Abstract: There are several issues to be considered to reduce the risk of rejection and minimize side effects associated with liver cell transplantation in chronic liver diseases. The source and the condition of stem cell proliferation and differentiation ex vivo and the transplantation protocols are important safety considerations for cell based therapy. The biochemical and molecular markers are important tools for safety evaluation of different processes of cell expansion and transplantation. Studies show that hepatocytes differentiated from adult and embryonic stem cells exhibit biochemical and metabolic properties resembling mature hepatocytes. Therefore these assays can help to assess the biological and metabolic performance of hepatocytes and progenitor stem cells. The assays also help in testing the contribution of transplanted hepatocytes in improving the repair and function of damaged liver in the recipient. Here we review the biochemical and metabolic markers, which are implicated in evaluation of safety issues of stem cells used for therapeutic purposes in chronic liver diseases and regeneration of damaged liver. We also highlight application of biochemical tests for assessment of liver cell transplantation. [Copyright &y& Elsevier]

Abstract: This study compared the sensitivity of differentiated hepatocyte-like cells, their progenitor mesenchymal stem cells (MSCs) and CD34+ stem cells to DNA damage and toxicity induced by aflatoxin B1 (AFB1). The hepatocyte-like cells and their progenitor cells (isolated from umbilical cord blood (UCB)) were each treated with AFB1 on day 15 of differentiation. Cell toxicity and genotoxicity effects were assessed using MTT and alkaline comet assays. AFB1 treatment resulted in a dose- and time-dependent inhibition of cell growth. The IC50 values of AFB1 for hepatocytes differentiated from CD34+ and MSCs were within the same range (44.7–46.8μM). The IC50 calculated for non-differentiated MSCs and CD34+ cells was slightly lower (42.0–43.4μM) than that calculated for their differentiated counterparts. However, the extent of DNA damage was different in differentiated and non-differentiated cells. The percentages of DNA (% DNA) in comet tails measured in hepatocytes differentiated from MSCs exposed to AFB1 (0, 2.5, 10 and 20μM) for 24h were ∼15, 55, 65 and 70%, respectively. In comparison, hepatocytes from CD34+ cells were more resistant to AFB1-induced DNA damage. Hepatocyte-MSCs were most sensitive to DNA damage, followed by UCB-CD34+ cells, then UCB-MSCs and finally hepatocyte-CD34+ cells. These results clearly showed that stem cells from different sources have different sensitivities to DNA damaging agents. These differences can be assigned to the expression levels of cytochrome P450 (CYP) particularly CYP3A4 in non-differentiated and differentiated cells. These data are useful in better understanding the susceptibility/resistance of stem cells in the process of differentiation to environmental toxicants. [Copyright &y& Elsevier]

Carcinoma antigen 125 (CA 125) is overexpressed in ovarian cancer and antibodies against it are widely employed for diagnostic purposes. The rarity of CA 125 antigenic domains and its highly glycosylated structure, however, is a problem that may prevent immunized mice from developing a diversified population of anti-CA 125 antibodies. In this study a prime-boost strategy, which potentially could augment the humoral immune responses against rare and poorly immunogenic determinants, was used for immunization of mice and monoclonal antibodies (mAbs) were produced by hybridoma technology. Reactivity of mAbs was then assessed by ELISA, western blotting, immunoprecipitation, immunohistochemistry and immunofluorescence staining of OVCAR-3 cell line. Altogether, 10 clones were produced, 3 of which had IgG isotype and the rest were IgM. Two-third of clones recognized cognate antigen in fixed and living cells and had strong immunoreactivity in IHC staining. In Western blotting, our antibodies recognized CA 125 as high molecular weight antigen mostly migrated in the 3% stacking gel. Immunoprecipitation of OVCAR-3 cell lysate by mAbs resulted in a very similar migration pattern that reconfirmed their specificities. The mAbs produced in this study are invaluable tools in diagnosis and research fields for assessment of CA 125 expression in cancerous ovarian tissues. [ABSTRACT FROM AUTHOR]

Abstract: Background: The molecular and cellular mechanisms linking chronic inflammation and gastrointestinal malignancy are not known with certainty. Aim: To investigate changes in potential causative factors during progression of esophagus cancer in a population living in high-risk area in Iran. Subjects: Formalin-fixed, paraffin-embedded esophageal specimens (n =87) from patients with gastroesophageal reflux disease (GERD), Barrett''s metaplasia, adenocarcinoma (ADC) and squamous cells carcinoma (SCC) were collected based on their pathological diagnosis. Methods: Immunohistochemical (IHC) technique was used to study tissue accumulation of P53, P21, cyclooxygenase-2 (COX-2), glutathione S-transferase-P (GST-Pi) and nitrotyrosine (NT) in patients and controls. Results: P53 expression was not detected in esophageal tissues from normal and GERD samples, whereas it was found positive in Barrett''s, ADC, and SCC samples. P21 positive sample was relatively higher in ADC patients as compared to that in SCC (ADC: 52.6%; SCC: 25%). GST-Pi expression was equally accumulated in all the samples. NT was predominantly expressed in ADC (72.7%). COX-2 expression was significantly higher in Barrett''s (60.0%) and ADC (66.6%) as compared to that in GERD, SCC and normal. These data were further confirmed by detecting the scores of immunostainings in all the positive samples. Conclusion: The pathological changes in ADC and SCC samples which were associated with increasing frequency of NT and COX-2 provides further evidence for involvement of these inflammatory factors in progression of esophagus cancer. [Copyright &y& Elsevier]

Abstract: The aim of this study was to find out the profile of cellular glutathione (GSH) and GSH S-transferase (GST) in hepatocytes differentiated from adult mesenchymal stem cells (MSC). For this purpose, we have derived functionally active hepatocyte-like cells from normal human multipotent adult MSC. Then the differentiated cells were characterized by specific hepatic markers. The cellular GSH and GST catalytic activity toward 1-chloro-2,4-dinitrobenzene (CDNB) were determined in hepatocyte-like cells differentiated from MSC compared with undifferentiated MSC. Reverse transcription polymerase chain reaction (RT-PCR) and immunoblotting techniques were used to study GST-P1-1 and GST-A1-1 expression in differentiated and undifferentiated cells. The results showed that there is more than threefold increase in GST catalytic activity in hepatocytes recovered by day 14 of differentiation. GST-P1-1 mRNA expression was detected in both differentiated hepatocyte-like cells and their undifferentiated progenitors. Under similar conditions, only differentiated hepatocyte-like cells expressed GST-A1-1 mRNA. These results were further confirmed by showing that the undifferentiated cells expressed both GST-A and GST-P proteins. Unlike GST, the level of cellular GSH was declined (∼20%) in hepatocytes derived from MSC as compared to that of undifferentiated cells. These data may suggest that hepatogenic differentiation of human bone marrow MSC is accompanied with the regulation of factors participating in GSH conjugation pathway. [Copyright &y& Elsevier]

Background: There is significant interest in using nanofibers in tissue engineering from stem cells. The transdifferentiation of mesenchymal stem cells into the hepatic lineage in a nanofibrous structure has not been reported. In this study, a three dimensional nanofibrous scaffold is introduced for differentiation of human bone marrow derived mesenchymal stem cells (hBMSCs) into hepatocytes. Methods: A scaffold composed of Poly (ε-caprolactone), collagen and polyethersulfone was fabricated by the electrospinning technique. After characterization of isolated hBMSCs, the performance of the cells on the scaffold was evaluated by Scanning Electron Microscopy (SEM) and MTT assay. Cytological, molecular and biochemical markers were measured to confirm differentiation potential of hBMSCs into hepatocytes. Results: The isolated cells possessed the basic properties of mesenchymal stem cells (MSCs). Based on scanning electron microscope (SEM) analysis and MTT assay, it was shown that the cells adhere, penetrate and proliferate on the nanofibers. Cultured cells on the nanofibers differentiated into hepatocyte-like cells and expressed hepatocyte specific markers such as albumin, α-fetoprotein, cytokeratin-18, cytokeratin-19 and cytochrome P450 3A4 at mRNA levels. Appearance of a considerable number of albumin-positive cells cultivated on the scaffold (47 ± 4%) as compared to the two-dimensional culture system (28 ± 6%) indicates the supporting role of the scaffold. The efficiency of the cells to produce albumin, urea, transferrin, serum glutamic pyruvic transaminase and serum oxaloacetate aminotransferase in hepatocytes on the scaffold further attest to the functionality of the cells. Conclusion: The data presented in this study show that the engineered nanofibrous scaffold is a conductive matrix which supports and enhances MSC development into functional hepatocyte-like cells. [ABSTRACT FROM AUTHOR]

Abstract: In this study characterization of endothelial cells differentiated from human bone marrow mesenchymal stem cells (hBMCs) was investigated in relation to their capillary network formation potential. Differentiation was performed in presence of vascular endothelial growth factor (VEGF) and insulin like growth factor-1 (IGF-1). A panel of cellular and molecular markers was used for characterization of the endothelial cells. The cells were strongly positive for von Willebrand factor (vWF) and vascular endothelial growth factor receptor 2 (VEGFR2) when measured at protein and mRNA levels. Development of endothelial cells was found to be associated with formation of typical organelles such as Weibel Palade (WP) bodies, Cavealae and pinocytic vesicles. Early vessel growth was also evidenced by showing specific junctions between the cells. The migratory and angiogenic properties of the cells were confirmed by showing capillary network formation in vitro. These results indicate that the capacity of endothelial cells differentiated from hBMSCs in formation of vascular system is consistent with molecular and structural development. [Copyright &y& Elsevier]

Abstract: Recently we reported that antioxidant system in brain and spinal cord in experimental autoimmune encephalomyelitis (EAE) mice is mainly affected at early stages of the disease [M. Zargari, A. Allameh, M.H. Sanati, T. Tiraihi, S.H. Lavasani, O. Emadyan, Relationship between the clinical scoring and demyelination in central nervous system with total antioxidant capacity of plasma during experimental autoimmune encephalomyelitis development in mice, Neurosci. Lett. 412 (2007), 24–28]. The aim of the present study was to investigate the role of uric acid (UA) on antioxidant system in liver and plasma of EAE mice. EAE was induced in C57/BL6 mice (n =60), followed by i.p. administration of UA (10mg/kg BW) in 30 mice at three distinct clinical stages (A: prior to onset, B: after onset, C: after development of EAE). Livers were removed and processed for measurement of lipid peroxidation products, reduced glutathione (GSH), and glutathione S-transferase (GST) and total antioxidant capacity of plasma (FRAP). The results showed that lipid peroxidation products in liver of EAE mice was increased significantly (∼85%) as compared to normal. UA administration to EAE mice caused a significant suppression of liver lipid peroxidation products (∼45%) at early stages (A and B). There was an inverse relationship between lipid peroxidation and cellular GSH in liver. GSH was significantly depleted in mice liver during the EAE progression, but it was recovered (∼29%) when UA was injected before the onset of the disease (groups A and B). Plasma total antioxidant capacity was significantly decreased during the development of EAE, however it was subsided in mice treated with UA as compared to the corresponding controls (21%) in groups A and B. Elevated liver GST as a result of EAE induction was reversed in mice treated with UA particularly in groups A and B. These results indicate that hepatic glutathione system, particularly GST plays a major role in modulation of oxidative damages to central nervous system (CNS) during EAE induction. The positive response of antioxidant system to UA administration in EAE mice was corroborated with improvement of clinical manifestation of the animals. [Copyright &y& Elsevier]

Abstract: Objectives: To investigate the interaction between welding fumes and noise in causation of hearing impairment. Methods: Groups of rabbits (n =6) were exposed to noise, welding fumes or combination of both prior to Distortion Product Otoacoustic-Emissions (DPOAEs) analysis. The function of outer hair cells (OHCs) was examined by DPOAE assessment over a broad range of frequencies. Variations in DPOAE amplitude were compared between control (n =6) and exposed (n =18) groups. Results: The DPOAEs levels measured at different frequencies (1379–6299Hz) were found to decrease significantly (P <0.05) in rabbits exposed to 110dB sound pressure level (SPL) broadband noise (8h/day, 12 days). In rabbits, exposed to carbon–steel welding fumes alone (157mg/m3), the threshold shift was limited to the high frequencies (2759–6299Hz), whereas, mixed exposure to noise and fumes resulted in reduction of DPOAEs at all the frequencies. Changes in DPOAEs were associated with increased susceptibility of erythrocytes to oxidation (P <0.05). Exposure to noise or fumes alone or simultaneously, suppressed total antioxidant ability of plasma as measured by ferric reducing ability of plasma (FRAP). Noise alone or in combination with fumes resulted in depletion of blood glutathione (GSH). Despite suppression of FRAP in the exposed groups, GSH was found to remain unchanged due to welding fumes suggesting that antioxidants other than GSH are affected by toxicants present in metal welding fumes. Conclusion: Exposure to very high levels of welding fumes can increase noise-related effects on OHC function by extending hearing threshold shift to wide band frequencies. [Copyright &y& Elsevier]

Abstract: Experimental autoimmune encephalomyelitis (EAE) was induced in a mouse model (C57/BL6) to investigate the antioxidant status of animals at various clinical stages of the disease. For this purpose, blood, brain and spinal cord samples from EAE mice were collected and examined at different scores following post-immunization with myelin oligodendrocyte glycoprotein (MOG). The clinical sign of mobility of animals on different days was associated with gradual increase in lipid peroxidation products (malondialdehyde, i.e. MDA) in brain and spinal cord. Changes in lipid peroxidation during EAE progression was inversely related to superoxide dismutase (SOD) activity in erythrocyte preparation. However, suppression of catalase in erythrocytes, tissue glutathione (GSH) and plasma total antioxidant capacity (FRAP assay) were the early events in EAE, occurred during scores 1 and 2. Biochemical alterations were corroborated with histopathological observations showing demyelination and inflammatory foci in central nervous system (CNS) of animals suffering from partial hind limb paralysis (score 3). These data suggest that generation of MDA in CNS is a continuous process during EAE induction and suppression of antioxidant factors are early events of the disease, but crucial in increasing the vulnerability of CNS to demyelinating lesions. [Copyright &y& Elsevier]

The effect of aflatoxin B1 (AFB1) on the expression of glutathione S-transferase-P (GST-P) which is the major isoform of GST in developmental stages has been investigated in rat liver during prenatal and postnatal stages. Following administration of AFB1 (0, 0.5, 1.0, 2.0, 3.0 or 4.0 mg/kg bw) injected I.P on day 8.5 of gestation the number of dead or reabsorbed fetuses and malformed embryos were recorded. Then the fetal livers were processed for measurement of total GST and GST-P activities, using 1-chloro-2,4-dinitrobenzene (CDNB) and ethacrynic acid (ETA) as substrates respectively. RT-PCR using rat GST-P specific primers was performed on mRNA extracted from livers. Besides, the effects of AFB1 on hepatic GST and GST-P were assessed in groups of suckling rats directly injected with the toxin. The results show that a single dose of AFB1 (1.0 or 2.0 mg/kg bw) caused approximately 50–60% depletion in fetal liver GST towards CDNB. Postnatal experiments revealed that liver GST (using CDNB as substrate) was significantly induced (~40%) in suckling rats injected with a single dose of AFB1 (3.0 mg AFB1/kg) 24 h before killing. Liver GST-P expression was unaffected due to AFB1 exposures of rats before and after the birth. This finding was substantiated by western blotting and RT-PCR techniques. These data suggest that AFB1-related induction in rat liver total GST after birth may be implicated in protective mechanisms against AFB1. In contrast, inhibition of this enzyme in fetal liver following placental transfer of the carcinogen may explain high susceptibility of fetal cells to transplancental aflatoxins. Furthermore, lack of influence of AFB1 on GST-P expression in developmental stages can role out the involvement of this class of GST in AFB1 biotransformation. [ABSTRACT FROM AUTHOR]

Abstract: Aflatoxicosis is one of the major causes of mortality and low productivity in broiler chickens, which is induced by aflatoxin contaminated grains. Elimination of aflatoxins by treatment with ammonia is one of the approaches to reduce aflatoxicosis. This study was conducted to assess the production parameters with respect to biochemical and histopathological changes in chicks fed ammonia treated maize. Detoxification of aflatoxin contaminated maize grains was done in a pilot plant with aqueous ammonia (1%, v/w). One-day-old broiler chicks (Ross 308) were fed on different dietary treatments designed dividing 480 chicks into six groups. Various dietary treatments included: group A (control), basal diet containing uncontaminated maize; group B, basal diet containing ammonia treated uncontaminated maize; group C, basal diet containing aflatoxin B1 (AFB, 1ppm); group D, basal diet having ammonia treated aflatoxin contaminated (1ppm) maize; group E, basal diet containing AFB (2ppm) contaminated maize; group F, basal diet containing ammonia treated AFB (2ppm) contaminated maize. Chickens were monitored daily and then body weight and feed consumption were recorded. At the end of rearing period (6 weeks), aflatoxin treated groups resulted in significant (P &#60;0.05) increase in mortality rate as compared to the dietary treatments having either uncontaminated and/or ammonia treated grains. Plasma enzymatic and non-enzymatic parameters showed progressive changes in birds examined 21 or 42 days after dietary aflatoxin treatments. In contrast to non-enzymatic parameters, marker enzymes particularly lactate dehydrogenase (LDH) and aspartate amino transferase (AST) showed more dependency on the concentration and duration of the exposure to aflatoxins. There was no significant change in dietary intake, body weight gain, and feed conversion ratio in chickens fed ammonia treated aflatoxin contaminated maize, whereas these parameters were suppressed in birds fed aflatoxin-containing diet. These data suggest that replacement of aflatoxin-containing maize with ammoniated grains can significantly suppress aflatoxicosis, leading to improvement in production parameters in broilers. [Copyright &y& Elsevier]

Objective Materials and Methods Results Conclusion Fibroblast growth factor receptor‐2 (FGFR2) and miR‐889‐3p expression in oral squamous cell carcinoma (OSCC) tumours were compared to normal controls. We then examined the relationship between miR‐889‐3p, FGFR2 expression and patient clinicopathological features.The interaction of FGFR2 and miR‐889‐3p was investigated using bioinformatics. Then, OSCC tumour biopsies and normal gingiva were collected and processed for expression analysis of FGFR2‐specific mRNA and miR‐889‐3p using real‐time PCR. Immunohistochemistry evaluated the expression of the FGFR2 protein.The protein and mRNA expression levels of FGFR2 were significantly greater in tumours when contrasted with controls. Expression of miR‐889‐3p was significantly lower in OSCC compared to normal tissues. The FGFR2 and miR‐889‐3p expressions were inversely related (−0.86 and −0.73, respectively) in both cases and controls. Changes in miR‐889‐3p and FGFR2 expression in tumour tissues were associated with lymph node metastasis (LNM), with ~0.57 and ~3.0 folds of change in positive‐LNM patients, respectively.Decreased expression of miR‐889‐3p in OSCC tumours suggests that miR‐889‐3p functions as a tumour suppressor gene. Overexpression of FGFR2 further proves the role of miR‐889‐3p in the regulation of the FGFR2 pathway. This was further confirmed by showing differences in miR‐889‐3p expression in positive and negative LNM cases. [ABSTRACT FROM AUTHOR]

Simple Summary: This manuscript focuses on the complex relationships between autophagy and epithelial mesenchymal transition (EMT) in cancer. Autophagy, a cellular degradation process, and EMT, a mechanism where epithelial cells acquire mesenchymal features, both play significant roles in cancer development. This review aims to explore how these processes interact, particularly how autophagy impacts cancer cell fate during EMT. The findings from this study are expected to contribute to a better understanding of cancer biology and could potentially impact cancer treatment strategies, as both autophagy and EMT are considered targets for therapy. Epithelial Mesenchymal Transition (EMT) is a dedifferentiation process implicated in many physio-pathological conditions including tumor transformation. EMT is regulated by several extracellular mediators and under certain conditions it can be reversible. Autophagy is a conserved catabolic process in which intracellular components such as protein/DNA aggregates and abnormal organelles are degraded in specific lysosomes. In cancer, autophagy plays a controversial role, acting in different conditions as both a tumor suppressor and a tumor-promoting mechanism. Experimental evidence shows that deep interrelations exist between EMT and autophagy-related pathways. Although this interplay has already been analyzed in previous studies, understanding mechanisms and the translational implications of autophagy/EMT need further study. The role of autophagy in EMT is not limited to morphological changes, but activation of autophagy could be important to DNA repair/damage system, cell adhesion molecules, and cell proliferation and differentiation processes. Based on this, both autophagy and EMT and related pathways are now considered as targets for cancer therapy. In this review article, the contribution of autophagy to EMT and progression of cancer is discussed. This article also describes the multiple connections between EMT and autophagy and their implication in cancer treatment. [ABSTRACT FROM AUTHOR]

Background: Oral Squamous Cell Carcinoma (OSCC) is among the ten most common cancers worldwide. Hypermethylation of CpG sites in the promoter region and subsequent down-regulation of a tumor suppressor gene, TGM-3 has been proposed to be linked to different types of human cancers including OSCC. In this study, methylation status of CpG sites in the promoter region of TGM-3 has been evaluated in a cohort of patients with OSCC compared to normal controls. Methods: Forty fresh tissue samples were obtained from newly diagnosed OSCC patients and normal individuals referred to dentistry clinic for tooth extraction. DNA was extracted, bisulfite conversion was performed and it was subjected to PCR using bisulfite-sequencing PCR (BSP) primers. Prepared samples were sequenced on a DNA analyzer with both forward and reverse primers of the region of interest. The peak height values of cytosine and thymine were calculated and methylation levels for each CpG site within the DNA sequence was quantified. Results: Quantitative DNA methylation analyses in CpG islands revealed that it was significantly higher in OSCC patients compared to controls. DNA methylation at CpGl/CpG3/CpG5 (p=0.004-0.01) and CpGl/CpG3 (p=0.001-0.019) sites was associated with tumor stage and grade, respectively. Male OsCC patients had higher methylation rate at CpG3 (p=0.032), while smoker patients showed higher methylation rate at CpG6 (p=0.045). Conclusion: These results manifested the contribution of DNA methylation of TGM-3 in OSCC and its potential association with clinico-pathologic parameters in OSCC. [ABSTRACT FROM AUTHOR]

The association of haptoglobin (Hp) with various cancers has been reported and also it has been documented that the Hp phenotypes/genotypes have different functional ability. So, we examined phenotypes/genotypes of Hp in newly diagnosed, untreated non-muscle invasive bladder cancer (NMIBC) patients and investigated its prognostic value for risk stratification of the cancer. In eighty NMIBC patients and 80 healthy individuals the Hp genotypes and phenotypes were analyzed using polymerase chain reaction (PCR) and two-dimensional gel electrophoresis (2D-GE), respectively. Besides, the presence of the Hpα1, α2, and β chains in the sera was confirmed by Mass Spectrometry (MS). The frequencies of the 1-1 and 2-2 genotypes/phenotypes were respectively higher and lower in healthy subjects compared to the patients. Our results revealed that the 2-2 genotype/phenotype could increase the risk of NMIBC. There was a positive association between the 2-2 genotype/phenotype with the T category/grade of cancer (p<0.05). The present study implied a strong association between the Hp phenotypes and genotypes with NMIBC. It was found that the 2-2 genotype and phenotype could be a risk factor for NMIBC incidence, as well as, progression. This study introduced Hp genotyping as a possible cost-effective and precise method for prognosis of individuals at the risk of NMIBC. [ABSTRACT FROM AUTHOR]

Abstract The anti-inflammatory role of the active metabolite of vitamin D, namely 1, 25-dihydroxyvitamin D 3 (calcitriol), has been reported in multiple sclerosis (MS). Moreover, recent studies have shown that fibroblast growth factor 23 (FGF23) is involved in the regulation of calcitriol biosynthesis. The probable changes of FGF23 and calcitriol concentrations in the CSF and serum of patients with MS were evaluated. Calcitriol concentration in the CSF and serum of MS patients was significantly higher than that in non-MS patients, while FGF23 concentration in MS patients was comparable to controls. We concluded that calcitriol concentration increases in the CSF and serum of MS patients independent of FGF23 status. Graphical abstract Unlabelled Image Highlights • Calcitriol concentration increases in the CSF and Serum of MS patients. • FGF23 concentration in CSF and Serum of MS patients is unchanged. • Calcitriol alters in the CSF and serum of MS patients independent of FGF23. • Calcitriol and FGF23 changes in CSF of MS patients are inversely related to EDSS. [ABSTRACT FROM AUTHOR]

Abstract Growing broiler chicks on a diet contaminated with aflatoxins (200 or 2000 ppb) for entire growth period resulted in increased oxidative stress and liver damage markers. The toxicity was subsided in broilers received a specific aflatoxin-binding probiotic i.e., Lactobacillus plantarum 299v (Lp299v). There was a substantial (30–90%) increase in antioxidant activity of plasma, which was suppressed due to dietary aflatoxins. Probiotic also reduced serum lactate dehydrogenase and alanine amino transferase together with lipid peroxidation products in liver, which were elevated due to aflatoxin. Because of Lp299v consumption, there was ∼20–55% recovery in body weight gain in broilers intoxicated with aflatoxins. Comparison of the Lp299v effects with that of a commercial aflatoxin binder revealed that, improved antioxidant activity of the chicks was associated with growth performance. These data suggest that aflatoxin-binding probiotics are beneficial with multi-functional effects and can efficiently help reducing aflatoxins in food chain associated with poultry industry. Highlights • L. plantarum 299v supplemented via drinking water protects chickens against aflatoxin-induced hepatotoxicity. • Chickens on drinkable L. plantarum 299v are benefited from improved total antioxidant activity. • The drinkable L. plantarum 299v in drinking water improves performance and growth in chickens. [ABSTRACT FROM AUTHOR]

Introduction: High dose of acetaminophen (APAP) is known to have hepatotoxic and nephrotoxic effects and studies show that this toxicities are dependent on the function of phase I bioactivation enzymes- Cyp450- and phase II biotransformation enzymes especially glucuronosylation and sulfonation pathways. However, the role of cyclooxygenase (COX) as an inflammatory mediator in toxic effects of APAP has not been explained satisfactorily yet. Methods: In this study, we aimed to find out if there is any association between APAP hepatotoxicity and COX-2 expression at mRNA levels. Male Balb/C mice were treated with a single high dose (300 mg/kg BW) or low dose (30 mg/kg BW) of APAP. Results: Following APAP treatment, serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) enzymes were measured as the biochemical markers of hepatocellular damage. Then liver and kidney biopsies were processed and examined for histolopathogical changes as well as for total RNA extraction and COX-2 gene expression. Serum ALT/AST levels were significantly (P<0.05) higher and there were hepatotoxic damages after 24 hours in mice exposed to high dose of APAP (300 mg/kg BW). However, no obvious nephrotoxicity was observed in mice treated with either low or high doses of APAP. Based on RT-PCR data, the COX-2 specific mRNA was not expressed in liver tissues of either control or APAP-treated mice, while, it was expressed in kidney tissues of both control and APAP-treated mice. Conclusion: These data may suggest that unlike in liver, COX-2 expression in kidney may play a protective role in APAP-related hepatotoxicity. [ABSTRACT FROM AUTHOR]

Klotho, as an antiaging protein, is involved in the maintenance and differentiation of neuronal or glial cells and, therefore, has been noticed as a potential therapeutic target for neurodegenerative disorders. Expression of Klotho has been examined in different cells and organs, however, our information about the developmental pattern of this protein during differentiation of mesenchymal stem cells (MSCs) into neuron‐like cells is limited. In this study, we conducted neural differentiation of mouse bone marrow‐derived‐MSCs and monitored the expression of Klotho together with selected neuron‐specific genes at messenger RNA (mRNA) on days 7 and 14 of differentiation using quantitative real‐time PCR. In addition, Klotho status at protein level was evaluated by immunocytochemistry. The results showed a significant change in the morphology of MSCs towards neuron‐like cells. These changes were observed with progressive growth and formation of cell connections towards the formation of a chain of neuron‐like cells which occurred in the second week of differentiation. Morphological changes were associated with a significant increase in the expression of neuron‐specific genes like pax‐6, neuN and, neurofilaments (NfL). Likewise, there was an increased expression of Klotho mRNA, and accumulation of Klotho protein in neuronal cell bodies, during the cellular differentiation of MSCs. These findings provided new evidence that neuronal differentiation from the MSCs is associated with increased expression of Klotho. These data may provide insight into the importance of Klotho protein in stem cell differentiation and regeneration in response to cell death in the central nervous system. Significance: Neuronal development is a complicated process. Specific growth factors have been identified to play role in this process. In this paper, we provide experimental evidences showing that in vitro differentiation of neuron cells from mouse bone marrow mesenchymal stem cells (MSCs) is associated with increased expression of Klotho, an anti‐ageing protein. This finding was confirmed by showing that morphological changes were associated with a significant increase in the expression of a panel of neuron‐specific genes Pax‐6, neuN, and, Nefl in the cells. During this process, klotho expression at mRNA level was increased, which was corroborated with klotho accumulation in cell bodies. These findings suggest that klotho contribute to neuronal cell differentiation and regeneration of these cells. [ABSTRACT FROM AUTHOR]

Background we recently showed that a hypothesized anti-aging and anti-inflammatory protein, namely Klotho, may contribute to the etiology and/or pathogenesis of multiple sclerosis (MS). In addition, Klotho function and its gene expression are dependent on inflammatory pathways. Accordingly, the aim of this study was to investigate the Klotho gene expression within peripheral blood mononuclear cells (PBMCs) of patients with MS. Methods Altogether, 30 patients with relapsing-remitting MS (RRMS) along with 30 age and sex-matched healthy individuals were enrolled in this study. Blood samples were obtained from all participants and then PBMCs were isolated. The quantitative Real-Time PCR was carried out for Klotho mRNA derived from PBMCs. Results The results showed that klotho gene expression in the PBMCs of patients with RRMS is nearly 2.5-fold less than healthy individuals ( P = 0.0006). Conclusion This is the first study demonstrating a possible role of Klotho in the PBMCs of MS patients. [ABSTRACT FROM AUTHOR]

Aflatoxin contamination of grains and agro-products is a serious food safety issue and a significant economic concern worldwide. In the present study, the effects of eugenol on Aspergillus parasiticus growth and aflatoxin production were studied in relation to the expression of some essential genes involved in aflatoxin biosynthetic pathway. The fungus was cultured in presence of serial two-fold concentrations of eugenol (15.62-500 μg mL) for 3 days at 28 °C. Mycelia dry weight was determined as an index of fungal growth, while aflatoxin production was assessed by high performance liquid chromatography. The expression of aflatoxin biosynthetic genes including ver- 1, nor- 1, pksA, omtA and aflR were evaluated by real-time PCR. Eugenol strongly inhibited A. parasiticus growth in the range of 19.16-95.83 % in a dose-dependent manner. Aflatoxin B production was also inhibited by the compound in the range of 15.07-98.0 %. The expressions of ver- 1, nor- 1, pksA, omtA and aflR genes were significantly suppressed by eugenol at concentrations of 62.5 and 125 μg mL. These results indicate that eugenol may be considered as a good candidate to control toxigenic fungal growth and the subsequent contamination of food, feed and agricultural commodities by carcinogenic aflatoxins. [ABSTRACT FROM AUTHOR]

Shift work is associated with sleep deprivation, occupational stress, and increased risk of depression. Depressed patients show increased oxidative stress. During excessive oxidative stress, Malondialdehyde (MDA) increases and total antioxidant capacity (TAC) decreases in body. This cross-sectional studywas conducted to determine the serumlevel of TACandMDAamong depressed rotational shift workers in Shahid Tondooyan Tehran Oil Refinery. 21-item Beck Depression Inventory was used to measure depression level. The level of TAC andMDA was measured by 8 mL fasting blood sample.MDA was determined by thiobarbituric acid reaction. Serum total antioxidants were measured using the ABTS. Results of this study showed that TAC mean and standard deviation concentration was 2.451 (±0.536)mg/dL and MDA was 3.725 (±1.098) mic.mol/L, and mean and standard deviation of depression score and BMI were 14.07 (±3.84) and 24.92 (±3.65) kg/m2, respectively. Depression score had a positive correlation with rotational shift work experience and work experience (r = 0.218 and r = 0.212), respectively, (p < 0.05). [ABSTRACT FROM AUTHOR]

Aflatoxins are secondary metabolites that are produced by toxigenic strains of some Aspergillus species on foods. Neem plant is a known inhibitor of aflatoxin production. We studied the effects of different concentrations of aqueous neem leaf extract on fungal growth and aflatoxin production by Aspergillus parasiticus (NRRL 2999) at different incubation times. The toxigenic fungus was cultured on sucrose low salts medium in the presence of various concentrations of extracts (0.2, 0.8, 3.12, 12.5 and 50% v/v). After shaking incubation of cultures for 2, 4, 6, 8, 10 and 12 days at 28 °C, the fungal mycelia was collected and processed for determination of dry weight. Mycelia and culture media were assayed by TLC method to detect aflatoxin B1 (AFB1). The extracts did not have any obvious effect on fungal growth. AFB1 production in the control samples increased to the maximum level on the 8th day. The inhibition of aflatoxin synthesis by plant extracts was found to be time- and dose-dependent. The maximum inhibitory effect was 80–90% in the presence of 50% concentration that when compared with control samples was significant ( P < 0.05). AFB1 secretion/production ratio in all of control and treated samples, other than 2nd day, approximately stayed and neem had no effect on it. [ABSTRACT FROM AUTHOR]