A systematic review on garbage enzymes and their applications in environmental processes.

Understanding the new insight on conversion of organic waste into value-added products can improve the environmental activities driven by microorganisms and return the nutrients to environment and earth. Here, we comprehensively review the available knowledge on application of garbage enzyme (GE) for different environmental activities including waste activated sludge, composting process, landfill leachate treatment, soil remediation and wastewater treatment with special focus on their efficiency. To identify peer-reviewed studies published in English-language journals, a comprehensive search was performed across multiple electronic databases including Scopus, Web of Science, Pubmed, and Embase. The search was conducted systematically using relevant keywords. The eligible studies were analyzed to extract data and information pertaining to components of GE, fermentation process operational parameters, type of hydrolytic enzymes and improved environmental performance. The findings derived from this current review demonstrated that GE produced from the fruit and vegetable peels, molasses or brown sugar (carbon source), and water within fermentation process contain different hydrolytic enzymes in order to facilitate the organic waste degradation. Therefore, GE can be considered as a promising and efficient pathway in order to improve the environmental activities depended on microorganism including, composting, wastewater and leachate treatment and bioremediation process.

The Expression Analysis of Long Non-coding RNAs Related to Wnt/ß-Catenin Signaling in Pancreatic Cancer Patients.

Background The oncogenic Wnt/ß-catenin signaling plays a critical role in carcinogenesis, prognosis, and resistance to therapy. Pancreatic cancer (PC) has high mortality because of its poor prognosis. Several studies have suggested that lncRNAs are directly involved in the development and progression of PC as well as in Wnt/ß-catenin signaling. In this study, we investigated and compared the expression of Wnt/ß-catenin signaling-related ZFAS1 and HCG11 lncRNAs, and their targets, CTNNB1 and IGF2BP1 genes in the blood of patients with PC and healthy individuals. A total of 47 PC patients and 50 healthy individuals participated in this study. RNA was extracted from the peripheral blood samples of participants, and cDNA was synthesized. The expression level of the selected genes was quantified by real-time PCR. The expression of HCG11 lncRNA and CTNNB1 genes in patients with PC was significantly upregulated compared to healthy individuals, and the expression of the ZFAS1 lncRNA was significantly downregulated. According to the analysis of the ROC curve, the diagnostic powers of ZFAS1 and CTNNB1 in PC were 0.67 and 0.69, respectively. Altogether, the present study suggests a role for ZFAS1 and HCG11 lncRNAs and CTNNB1 and IGF2BP1 in the pathogenesis of pancreatic cancer. Moreover, the peripheral expression of these lncRNAs may be useful as potential biomarkers for PC.

The oxidative stress-associated long non-coding RNAs in pancreatic cancer.

PURPOSE: A lot of people are dying from pancreatic cancer (PC) annually. The early detection of this cancer is particularly challenging due to the fact that symptoms tend to appear in advanced stages. It has been suggested that oxidative stress may play a role in the development of PC. Several genes regulate this process, including long noncoding RNAs (lncRNAs). There is no comprehensive study on the expression pattern of lncRNAs related to oxidative stress in PC patients. In the present case-control study, we quantified levels of oxidative stress-associated lncRNAs in PC patients versus healthy controls. PATIENTS AND METHODS: In the present study, we investigated the expression levels of lincRNA-p21, LUCAT, RMST, FOXD3-AS1, and MT1DP lncRNAs in the peripheral blood mononuclear cells (PBMCs) of 53 â€‹PC patients and 50 healthy controls. The association between lncRNA expression and clinical and pathological characteristics was also evaluated. RESULTS: The expression of lincRNA-P21 and rhabdomyosarcoma 2-associated transcript (RMST) lncRNAs in PC patients has significantly decreased. Expression of lncRNA RMST was significantly higher in TNM stage III-IV patients in comparison to TNM stage I-II patients. In addition, a significant positive association was recognized between candidate lncRNA expression, and finally, the AUC values of the expression levels of lincRNA-p21 and RMST were 0.60 and 0.61, respectively. CONCLUSIONS: Altogether, our study suggests a possible role of lincRNA-p21 and RMST lncRNAs in the etiology of PC pathobiology, and their biomarker role may be understood in future studies.

Antiproliferative and Antitelomerase Effects of Silymarin on Human Colorectal and Hepatocellular Carcinoma Cells.

Silymarin, a widely-used hepatoprotective agent, has shown antitumor properties in both in vitro and animal studies. Currently, there is limited knowledge regarding silymarin's antitelomerase effects on human colorectal cancer and hepatocyte carcinoma cells. In this study, we investigated the antiproliferative and antitelomerase effects of silymarin on four human colorectal cancer and HepG2 hepatocyte carcinoma cell lines. The cell viability and telomerase activity were assessed using MTT and the telomerase repeat amplification protocol assay, respectively. We also investigated the effects of silymarin on the expression of human telomerase reverse transcriptase and its promoter methylation in HepG2 cells by real-time RT-PCR and methylation-specific PCR, respectively. Silymarin treatment inhibited cell proliferation and telomerase activity in all cancer cells. After 24 h of treatment, silymarin exhibited IC50 values ranging from 19 - 56.3 µg/mL against these cancer cells. A 30-min treatment with silymarin at the IC50 concentration effectively inhibited telomerase activity in cell-free extracts of both colorectal cancer and hepatocyte carcinoma cells. Treatment of HepG2 cells with 10 and 30 µg/mL of silymarin for 48 h resulted in a decrease in human telomerase reverse transcriptase expression to 75 and 35% of the level observed in the untreated control (p < 0.01), respectively. Treatment with silymarin (10, 30, and 60 µg/mL) for 48 h did not affect human telomerase reverse transcriptase promoter methylation in HepG2 cells. In conclusion, our findings suggest that silymarin inhibits cancer cell growth by directly inhibiting telomerase activity and downregulating its human telomerase reverse transcriptase catalytic subunit. However, silymarin did not affect human telomerase reverse transcriptase promoter methylation at the concentrations of 10 - 60 µg/mL used in this study.

Angiopoietin-Like Proteins 2 and 3 in Children and Adolescents with Obesity and Their Relationship with Hypertension and Metabolic Syndrome.

BACKGROUND: Angiopoietin-like protein 2 (ANGPTL2) is one of the adipocyte-derived inflammatory factors which connects obesity to insulin resistance. ANGPTL3 has a direct role in regulation of lipid metabolism. The objective of this study was to evaluate ANGPTL2 and ANGPTL3 in childhood obesity and their relationship with metabolic syndrome. METHODS: 70 children and adolescents, 35 obese and 35 normal-weight subjects, were enrolled in this research after complete clinical examination and anthropometric evaluations. Serum ANGPTL2 and ANGPTL3 and insulin were measured by enzyme-linked immunosorbent assay (ELISA). Homeostatic model assessment of insulin resistance (HOMA-IR) was calculated and used to estimate insulin resistance (IR). Colorimetric methods were used for the assessment of fasting plasma glucose (FPG), LDL-C, HDL-C, total cholesterol (TC), and triglyceride (TG). RESULTS: The levels of ANGPTL2 and ANGPTL3 were significantly higher in obese subjects than those in controls, but they did not differ significantly in subjects with or without IR. ANGPTL3 was found to be significantly elevated in obese children with metabolic syndrome (MetS) in comparison with those without MetS. Both of the studied ANGPTLs were positively correlated with BMI, systolic blood pressure (SBP), diastolic blood pressure (DBP), TC, and LDL-C. The correlation between ANGPTL3 and either TC or LDL-C remained significant after adjusting for BMI. CONCLUSION: Serum ANGPTL2 and ANGPTL3 were elevated in obesity and associated with blood pressure and indices of metabolic syndrome, suggesting that they might be involved in the advancement of obesity-related hypertension and metabolic syndrome.

Nicotinamide Phosphoribosyltransferase Knockdown Leads to Lipid Accumulation in HepG2 Cells through The SIRT1-AMPK Pathway.

OBJECTIVE: Nicotinamide phosphoribosyltransferase (NAMPT), which is responsible for biosynthesis of nicotinamide adenine dinucleotide (NAD), has a regulatory role in cellular metabolism and thus, might be implicated in non-alcoholic fatty liver disease (NAFLD). This study aimed to show how NAMPT down-regulation in liver cells influences lipid metabolism and sirtiun 1 (SIRT1), as the main NAD-dependent deacetylase enzyme. MATERIALS AND METHODS: In this experimental study, HepG2 cells were transfected with NAMPT siRNA and hepatic triglyceride (TG) content and SIRT1 deacetylase activity were measured by colorimetric and fluorometric methods, respectively. Gene expression of fatty acid synthase (FAS) and sterol regulatory element-binding protein-1c (SREBP- 1c) was evaluated by real-time polymerase chain reaction (PCR). Total protein level and the phosphorylated form of acetyl-CoA carboxylase (ACC) and AMP-activated protein kinase (AMPK) were also investigated by western blotting. RESULTS: Knockdown of NAMPT significantly promoted the accumulation of TG in HepG2 cells, accompanied by a remarkable decline in SIRT1 deacetylase activity. A significant rise in the gene expression of two key lipogenic factors, FAS and SREBP-1c was also observed. These effects were also accompanied by decreased phosphorylation of ACC and AMPK. On the other hand, treatment of transfected cells with either NAD, as the SIRT1 substrate or resveratrol, as the SIRT1 activator reversed the outcomes. CONCLUSION: These results demonstrated a protective role for NAMPT against NAFLD and its involvement in the regulation of de novo lipogenesis through the SIRT1/AMPK pathway.

Effects of resveratrol on coagulative, fibrinolytic, and inflammatory marker expression and secretion by endothelial cells (human umbilical vein endothelial cells).

: Increasing the prevalence of cardiovascular disease (CVD) has led to an investigation into components that might influence CVD. Accordingly, many recent studies have reported the benefits of resveratrol (RSV). Therefore, this study aimed to scrutinize the direct effect of RSV on human umbilical vein endothelial cells (HUVECs) by detecting coagulative, fibrinolytic, and inflammatory markers. HUVECs were cultured and treated with different concentrations of RSV. The effects of RSV were identified by representative markers of coagulation, fibrinolysis pathway, and inflammation, including von Willebrand factor (VWF), factor VIII, tissue plasminogen activator-1 (t-PA-1), and interleukin-8 (IL-8). The detection process was carried out using real-time PCR (qPCR), flow cytometry, ELISA, and immunocytochemistry (ICC) methods. The present findings demonstrated a significant decrease in VWF, t-PA-1, and IL-8 secretion levels. Furthermore, RSV diminished the activity of factor VIII and mRNA expression levels of VWF and t-PA-1. The ICC results also showed a decrease in the level of intracellular t-PA. Our data revealed the anti-inflammatory, anticoagulation, and antifibrinolytic effect of RSV in cell culture.

The effect of trans-palmitoleic acid on cell viability and sirtuin 1 gene expression in hepatocytes and the activity of peroxisome-proliferator-activated receptor-alpha.

BACKGROUND: Accumulation of fatty acids in liver causes lipotoxicity which is followed by nonalcoholic fatty liver disease. The association between intakes of trans-fatty acids with metabolic diseases is still controversial. Accordingly, the objective of this study was to investigate the in vitro effects of trans-palmitoleic acid (tPA) and palmitic acid (PA) on lipid accumulation in hepatocytes, focusing on the gene expression of sirtuin 1 (SIRT1) as well as the transcriptional activity of peroxisome proliferator-activated receptor alpha (PPARα). MATERIALS AND METHODS: In this experimental study, hepatocellular carcinoma (HepG2) cells were cultured and treated with various concentrations of tPA and PA (C16:0). The accumulation of triglyceride in the cells was measured by enzymatic method. Gene expression was evaluated by real-time polymerase chain reaction. The activity of PPARα was assessed by luciferase reporter assay after transfection of human embryonic kidney 293T cells by a vector containing the PPAR response element. RESULTS: While concentration >1 mM for PA and cis-PA (cPA) reduced the viability of hepatocytes, tPA revealed an opposite effect and increased cell survival. Lipid accumulation in HepG2 cells after treatment with tPA was significantly lower than that in cells treated with PA. In addition, tPA at physiological concentration had no effect on the expression of SIRT1 while at high concentration significantly augmented its expression. There was a modest increase in PPARα activity at low concentration of tPA. CONCLUSION: tPA causes less lipid accumulation in hepatocytes with no detrimental effect on cell viability and might be beneficial for liver cells by the activation of SIRT1 and induction of PPARα activity.

Metformin reduces fibrosis factors in insulin resistant and hypertrophied adipocyte via integrin/ERK, collagen VI, apoptosis, and necrosis reduction.

AIMS: Fibrosis as the hallmark of adipose tissue dysfunction which is associated with insulin resistance and type 2 diabetes, results from deposition of excess extra cellular matrix components like collagen and increased cell death. Here we investigated the effect of antidiabetic drug, Metformin, on the factors that play role in fibrosis such as; integrin/ERK pathway, collagen VI, MMP2, MMP9, apoptosis markers including DAPK1, DAPK3, DAP, SIVA, necrosis markers including RIPK1, RIPK3, and MLKL in insulin resistant and hypertrophied adipocytes. METHODS: 3T3-L1 adipocytes after differentiation to insulin resistant and hypertrophied cells, treated with Metformin, and the gene expression of aforementioned factors assayed by real time PCR. The protein expression of collagen VI and ERK assayed by western blotting. KEY FINDINGS: The expression of integrins changed from 0.5 to 6-fold in hypertrophied adipocyte versus adipocyte. Apoptosis and necrosis markers increased >1.5-fold in insulin resistant and hypertrophied adipocytes. Also ECM components and ERK activation increased >2-fold and 1.5-fold, respectively in insulin resistant and hypertrophied adipocytes. Metformin caused reduction of activity of integrin/ERK pathway in Metformin treated insulin resistant and hypertrophied adipocytes compared to untreated group. Metformin also reduced collagen VI in both gene and protein expression level, MMP2 and MMP9 in gene expression, and also the expression of apoptosis and necrosis gene. SIGNIFICANCE: Metformin with reduction of ECM component as collagen VI, MMP2 and MMP9, integrin/ERK pathway, necrosis markers as RIPK1, RIPK3 and MLKL, and apoptosis markers including DAP, DAPK1, DAPK3 and SIVA effects on fibrosis in insulin resistant and hypertrophied adipocytes in vitro.

Plasma levels of CTRP-3, CTRP-9 and apelin in women with multiple sclerosis.

BACKGROUND: Recently, adipocytokines have been shown to play a pivotal role in autoimmune and inflammatory-related disease. The purpose of this study was to compare the levels of CTRP3, CTRP9, adiponectin and apelin- in Multiple Sclerosis (MS) patients with healthy subjects and their relationship with clinical parameters and the levels of pro-inflammatory mediators. METHODS: Plasma levels of CTRP3, CTRP9, apelin, TNF-α, hs-CRP, and adiponectin were evaluated in 24 healthy women and 26 women with relapsing-remitting MS using immunoassay methods. RESULTS: The plasma apelin level of the MS patients was significantly lower than that of healthy controls. The concentration of TNF-α and adiponectin were significantly higher in MS patients compared to the healthy controls. Plasma CTRP3, CTRP9 and hs-CRP levels were not significantly different between the two groups. There was no correlation between these adipokines and inflammatory mediators. A statistically significant negative correlation was observed between plasma concentrations of apelin with expanded disability status scale (EDSS) scores and number of relapse. CONCLUSIONS: Our findings suggest that adipokines, particularly apelin and adiponectin, may contribute to the pathogenesis of MS and can be considered as a biomarker or as a therapeutic target for the treatment of this disease.

The Generation of Hematopoietic Progenitor Cells from Endothelial Cells by Silymarin.

Background: The generation of hematopoietic cells from endothelial cells may determine a novel approach for the production of blood cells. Regenerative functions of Silymarin, a mixture of polyphenolic flavonoids derived from milk thistle, have previously been reported. The purpose of this study was to investigate the direct effect of Silymarin on conversion of endothelial cells to hematopoietic cells. Methods: The effect of Silymarin on CD34, CD45, and vascular endothelial growth factor receptor-2 (VEGFR-2) markers along with all the main hematopoietic lineage markers in human umbilical vein endothelial cells (HUVEC) were investigated by flow cytometry and immunocytochemistry. Results: Treatment of HUVEC cells with Silymarin after 24 hours significantly increased the number of the cells expressing CD34 and CD45 markers. Although the percentage of VEGFR-2 and CD13 was increased, the changes were not statistically significant during 24 hours. No significant expression of the rest of the hematopoietic lineage specific markers was found during this time. Conclusions: Taken together, our preliminary findings demonstrated the potential of Silymarin to switch human umbilical vein endothelial cells into hematopoietic progenitor cells.

Proliferative and Anti-Inflammatory Effects of Resveratrol and Silymarin on Human Gingival Fibroblasts: A View to the Future.

OBJECTIVES: It has been demonstrated that polyphenol components such as silymarin and resveratrol have anti-inflammatory properties. Periodontitis is a chronic inflammatory disease that leads to the breakdown of dental supporting tissues and tooth loss. The purpose of this study was to investigate the anti-inflammatory effects of silymarin and resveratrol on lipopolysaccharide (LPS)-induced inflammatory response in human gingival fibroblasts (HGFs). MATERIALS AND METHODS: HGFs were treated with different concentrations of silymarin and/or resveratrol (25, 50, 100 and 200µg/ml). The effects of silymarin and resveratrol on cell viability and proliferation were assessed by MTT assay and cell cycle analysis, respectively. Also, HGFs were treated with silymarin and/or resveratrol and were stimulated with LPS. The levels of Interleukin-6 (IL-6) and IL-8 were assessed by enzyme-linked immunosorbent assay (ELISA). RESULTS: After treatment with silymarin, the viability of fibroblasts significantly increased, whereas treatment with resveratrol did not have any significant effect on cell viability. However, the combination of these flavonoids (50µg/ml silymarin and 100µg/ml resveratrol) significantly increased the viability of fibroblasts. Resveratrol significantly inhibited LPS-induced IL-6 and IL-8 secretion by HGFs, but silymarin did not show such a significant effect. CONCLUSIONS: The findings of the present study demonstrated the anti-inflammatory effects of resveratrol and its combination with silymarin. Therefore, the combination of silymarin and resveratrol may be useful as a therapeutic agent for treatment of periodontal diseases.

Evaluation of Plasma TRB3 and Sestrin 2 Levels in Obese and Normal-Weight Children.

OBJECTIVE: Obesity in childhood and adolescence is associated with metabolic syndrome and cardiovascular diseases. TRB3 (Tribbles homolog 3) and sestrin 2 are two newly found proteins that have been identified to play an important role in obesity and its complications. AIM: The purpose of this study was to evaluate concentrations of TRB3 and sestrin 2 in plasma of obese and normal-weight children and adolescents, and their association with metabolic and anthropometric parameters. METHODS: Plasma levels of TRB3, sestrin 2, insulin, fasting plasma glucose (FPG), and lipid profile were evaluated in 70 children and adolescents (34 obese and 36 controls). Insulin resistance was calculated using a homeostasis model assessment of insulin resistance. Metabolic syndrome was defined according to IDF criteria. RESULTS: Plasma TRB3 levels of the obese subjects were significantly higher than that of normal weight subjects. TRB3 levels were positively correlated with BMI, BMI z-score, waist circumference, and FPG. The concentration of sestrin 2 was significantly lower in obese subjects compared to normal-weight subjects. A statistically significant positive correlation was observed between plasma concentrations of sestrin 2 and high-density lipoprotein cholesterol. Neither TRB3 nor sestrin 2 were correlated with insulin resistance and metabolic syndrome. CONCLUSION: Both TRB3 and sestrin 2 may contribute to the development of obesity and its complications and can be considered interesting therapeutic target for the treatment of obesity.

The effect of silymarin (Silybum marianum) on human skin fibroblasts in an in vitro wound healing model.

CONTEXT: Silymarin, a flavonolignan from Silybum marianum (L.) Gaertn. (Asteraceae), has been reported to have antioxidant and anti-inflammatory properties. Therefore, it may be worthwhile to study the effect of silymarin on wound healing. OBJECTIVE: To evaluate the effect of silymarin on human fibroblast cells in an in vitro model of wound healing. MATERIALS AND METHODS: Human fibroblast cells were treated with different concentrations (4.5, 9, 18, 36 µg/mL) of silymarin. The effects of silymarin on cell viability, proliferation, collagen synthesis, and expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthetase (iNOS) were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, 5-bromo-2'-deoxy-uridine, hydroxyproline analysis and real-time PCR, respectively. The effect of silymarin on cellular antioxidant status was determined by protection against hydrogen peroxide (H2O2)-induced cell injury and free radical scavenging activity (ABTS assay) of the cells. RESULTS: Results of the present study indicate that pretreatment of fibroblast cells with silymarin significantly protected cells against H2O2-induced injury (p < 0.05). After an 18 h treatment of cells with 36 µg/mL silymarin, total antioxidant capacity of cells significantly increased (p < 0.05). Furthermore, pretreatment of human fibroblast cells with silymarin significantly inhibited lipopolysaccharide (LPS)-induced COX-2 mRNA expression (p < 0.001). There was no significant difference in fibroblast proliferation and collagen synthesis between treatment and control groups (p > 0.05). DISCUSSION AND CONCLUSION: Silymarin may be useful as a therapeutic agent for the treatment of cutaneous wounds through its antioxidation and anti-inflammation effects.

Effect of topical application of silymarin (Silybum marianum) on excision wound healing in albino rats.

Silymarin, an extract from Silybum marianum, has been shown to have antioxidant properties. However, there is no scientific report on wound healing activity of the silymarin. The purpose of this study was to evaluate the effect of topical administration of silymarin on excision wound healing in rats. Excision wounds were made on the back of rats. Rats were divided into three groups, as control, vehicle, and treatment. Vehicle and treatment groups received polyethylene glycol and silymarin dissolved in polyethylene glycol, respectively. The control group did not receive any treatment. The wound tissues were removed on 5th, 10th and 15th day for histopathological analysis and total collagen determination by hydroxyproline assay. Results showed that silymarin increased epithelialization and decreased inflammation but did not have any effect on percentage of wound contraction, collagenization and hydroxyproline levels. It was concluded that silymarin can significantly stimulate epithelialization and reduce inflammation in full-thickness wounds in rats.

Effect of aqueous extract of Elaeagnus angustifolia fruit on experimental cutaneous wound healing in rats.

The present study was conducted to investigate the histological changes and wound healing effect of aqueous extract of Elaeagnus angustifolia. After creating full-thickness skin wounds on the back of 45 male Sprague-Dawley rats they were randomly divided into three groups. Treated group received the extract, positive control group were treated with mupirocin ointment 2% and control group did not receive any treatment. Wound healing rates were calculated on days 3, 5, 8, 10, 12 and 15 post-wounding and the wound tissues were harvested at 5, 10, and 15 days for histological analysis and hydroxyproline content measurement. The results indicated a significant increase in the percentage of wound contraction and hydroxyproline content in the treated group comparing to the control and positive control groups. A significant increase in the assigned histological scores was observed at 10 and 15 days in the treated and positive control groups compared to the control group. The results demonstrate that aqueous extract of Elaeagnus angustifolia accelerates cutaneous wound healing, and its effect may be due to the increased re-epithelialization and collagen deposition in wound and so it can be considered as a therapeutic agent for wound healing.

Immunohistochemical analysis of selected molecular markers in esophagus precancerous, adenocarcinoma and squamous cell carcinoma in Iranian subjects.

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.

Changes in hepatic cytosolic glutathione S-transferase activity and expression of its class-P during prenatal and postnatal period in rats treated with aflatoxin B1.

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 (approximately 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 trans-plancental 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.