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Stem cell therapy for degenerative diseases has been established; however there are controversies over the treatment of solid tumors with stem cell transplantation. In the present study, the anti-tumor action of mesenchymal stem cells (MSCs) has been examined in a mouse model of breast cancer with emphasize on tumor growth, angiogenesis and c-Myc expression in breast tumors. For this purpose, MSCs were isolated from bone marrow of Balb/c mice and characterized. A Balb/c mouse model of breast cancer was developed and subjected to cell therapy intra venous (I.V) or intra tumor (I.T) with MSCs. Tumor growth was measured using a digital caliber for until the end of experiment (30days). Then the mice were sacrificed and their tumors were removed and processed for histopathological examination, immunohistochemical assay of CD31 and measuring of c-Myc expression using quantitative PCR. Detection of the labeled-MSCs in tumors following injection of the cells (I.V or I.T) clearly showed the homing of MSCs into tumors. Tumor growth in case of MSC-treated mice by I.V and I.T routes was inhibited by approximately 28% and 34% respectively compared to controls. The suppression of angiogenesis was reflected in Micro Vessel Density (MVD) following I.V or I.T delivery of the MSCs. c-Myc gene expression in tumor tissues of mice treated I.V or IT with MSCs was down-regulated to 28.0% and 16.0% respectively compare to control groups. In conclusion, growth inhibition of breast tumors in mice due to MSC therapy is associated with modulation of c-Myc activation and angiogenesis markers.

Incorporation of endothelial cells or their progenitor cells into newly sprouting blood vessels can contribute to tissue vascularization after ischemic injury. However, the interaction of the stem cells-derived endothelial cells with angiogenesis within tumors is not well understood. The aim of this study was to examine the efficiency of endothelial-like cells derived from MSCs in controlling breast tumor growth associated with abnormal angiogenesis. For this purpose, Balb/c mouse model of breast carcinoma was developed and subjected to intra tumor (I.T)/intra venous (I.V) therapy with undifferentiated MSCs or endothelial cells derived from them. The homing of the stem cells was approved by measuring different markers as well as tracing green fluorescence protein (GFP)-labeled MSCs in the tumors. Tumor growth was measured following cell therapy using a digital caliper. At the end of treatment period (30 days) the angiogenesis markers; VEGFR2 expression as well as micro-vessel density (MVD) using CD31 were estimated in tumor tissues. Stem cell transplantation to mice bearing breast tumors resulted in tumor growth suppression in all experimental groups. The endothelial markers; CD31 and VEGFR2 were down regulated following I.T delivery of the endothelial cells. Accordingly, angiogenesis was suppressed following I.T administration of endothelial cells which was associated with increased focal necrosis in the tumors. In conclusion, data show that endothelial cells directly injected into tumors is more efficient compared to undifferentiated MSCs in controlling tumor-associated angiogenesis and tumor growth.

Abstract. Background: This study assessed the effect of supplementation of omega-3 fatty acids and ascorbic acid alone and in combination on the level of lipid peroxidation (malondialdehyde, MDA concentration), total antioxidant capacity (TAC) and depression scores in depressed rotational shift workers in the Tehran Shahid Tondgoyan Oil refinery. Material and methods: In this randomized, double-blind, placebo-controlled trial, 136 men who met the inclusion and exclusion criteria and had a Beck Depression Inventory (BDI) score ≥ 10 were selected. Among the participants, 33 received omega-3 fatty acid soft gel (1000 mg twice daily) with vitamin C (250 mg twice daily) (group 1), 31 took omega-3 fatty acid supplements and vitamin C placebo (group 2), 30 took omega-3 fatty acid supplement placebo and vitamin C (group 3), and 32 received omega-3 fatty acid supplement placebo and vitamin C placebo (group 4) for 2 months. Measured were serum MDA, TAC concentrations, and BDI scores at baseline and after 2 months. Results: This study showed that the BDI score was reduced significantly in all 4 groups, however, the level of decrease was more in the omega-3 fatty acid (alone) supplementation group (mean 6.29 score decrease) (p < 0.001). MDA level decreased significantly in groups with omega-3 fatty acids (mean 0.78 μmol/L ± 1.64 μmol/L decrease) (p = 0.014) or vitamin C supplementation alone (mean 0.74 μmol/L ± 1.55 μmol/L decrease) (p = 0.014), but not in combination. Conclusions: Supplementation of omega-3 fatty acids alone and not in combination with vitamin C had a better impact on depression and MDA level pronounced in depressed male shift workers.

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 cells in vitro via over-expressed LDLRs. The efficiency of the NPs uptake to malignant C6 glioma cells was remarkable compared to non-tumor glial cells (p p

Caraway (Carum carvi L.) is widely cultivated all over the world. It is among the widely used spices which are used as crude or essence in various food products for its pleasant flavor and antispoilage properties. The caraway seeds contain essential oils rich in nutraceutical compounds used as food supplements and plant-based medicine. Experimental evidences show that the essential oils (essence) and active ingredients such as carvone possess antioxidant and antimicrobal properties. Evidence shows that the essential oils are more effective when used as mixture suggesting the additive effects of the oil components. In this chapter, we introduce the caraway active components and their general applications particularly in food preservation and processing.

Two experiments were performed to screen bacilli isolated from quails for their aflatoxin removal potential and to assess the efficiency of their amelioration of experimental aflatoxicosis. Nonhemolytic bacilli were selected for in vitro aflatoxin B1 (AFB1) removal and conventional probiotic tests. The isolate with the highest scores was selected for assessment in field experiments and was identified as Berevibacillus laterosporus (Bl). In the second experiment, 125 male Japanese quails (21 d old) were divided into 5 groups with 5 replications to compare the toxin removal efficiency of Bl with that of a commercial toxin binder, improved Millbond-TX (IMTX). The experimental groups were as follows: Control (without any feed additive or AFB1); AFB1 (2.5 mg/kg); AFB1+Bl (2.5 mg/kg+10(8) cfu/mL); AFB1+IMTX (2.5 mg/kg+2.5 g/kg); and Bl (10(8) cfu/mL). The greatest BW gain and slaughter and carcass weights were found in the Bl group and the lowest values were observed in the AFB1 group (P

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 2.0 g/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 (2.0 g/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.

Telomerase has been proposed as a novel and potentially selective target in cancer therapy. Silymarin, which is a standardized mixture of flavonolignans from the medical plant Silybum marianum, has potent effects against various types of cancer cells, but its effect on telomerase activity in the human leukemia cell line K562 has not been investigated. The aim of this study was to examine the mechanism of silymarin-induced apoptosis in K562 cells, with particular emphasis on its effect on telomerase activity. The antiproliferation effect of silymarin on K562 cells was evaluated by the MTT assay. To measure apoptosis, Hoechst 33342 staining and flow cytometry were used. The telomerase activity was determined using the telomeric repeat amplification protocol (TRAP)-ELISA assay. The treatment of the K562 cells with silymarin resulted in a significant inhibition of cell growth and telomerase activity. Also, a positive correlation was found between telomerase inhibition and induction of apoptosis in silymarin-treated K562 cells. These results suggest a novel mechanism in the anticancer activity of silymarin in human leukemia K562 cells and may provide a basis for future development of anti-telomerase therapies.

We have recently reported that the inhibition of colonic premalignant lesions induced by 1,2-dimethylhydrazine (DMH) is mediated by the interference of caraway oil components in the activities of the main hepatic xenobiotic metabolizing enzymes. The present study was carried out to examine the effect of dietary caraway oils on the progression of cancer, with emphasis on β-catenin expression in the colon during DMH-induced colonic carcinogenesis. For this purpose, colon cancer was induced by DMH in rats (20 mg/kg body weight for 5 weeks) and groups of animals were given dietary caraway essential oils at two levels (0.01 and 0.1%) for 16 weeks. After 16 weeks and at the end of the experimental period the colon tissue biopsies were processed for histopathological examination and the expression of β-catenin at mRNA and protein levels was estimated by polymerase chain reaction and enzyme-linked immunosorbent assay. The formation of premalignant lesions based on aberrant crypt foci (ACF) in DMH-treated rats was greatly inhibited (72-87%) in rats given dietary essential oils when compared to respective controls. There was a correlation between the number of colonic ACF formation and the expression levels of β-catenin measured at protein and mRNA levels. These results indicate that the Wnt/β-catenin signaling pathway is activated during colon cancer promotion and that the expression of colonic β-catenin is altered in long-term caraway oil feeding, leading to suppression of DMH-induced premalignant lesions in rat colon.

In the present study, genetic diversity and mycotoxin profiles of Aspergillus flavus isolated from air (indoors and outdoors), levels (surfaces), and soils of five hospitals in Southwest Iran were examined. From a total of 146 Aspergillus colonies, 63 isolates were finally identified as A. flavus by a combination of colony morphology, microscopic criteria, and mycotoxin profiles. No Aspergillus parasiticus was isolated from examined samples. Chromatographic analyses of A. flavus isolates cultured on yeast extract-sucrose broth by tip culture method showed that approximately 10% and 45% of the isolates were able to produce aflatoxin B(1) (AFB(1)) and cyclopiazonic acid (CPA), respectively. Around 40% of the isolates produced sclerotia on Czapek-Dox agar. The isolates were classified into four chemotypes based on the ability to produce AF and CPA that majority of them (55.5%) belonged to chemotype IV comprising non-mycotoxigenic isolates. Random amplified polymorphic DNA (RAPD) profiles generated by a combination of four selected primers were used to assess genetic relatedness of 16 selected toxigenic and non-toxigenic isolates. The resulting dendrogram demonstrated the formation of two separate clusters for the A. flavus comprised both mycotoxigenic and non-toxigenic isolates in a random distribution. The obtained results in this study showed that RAPD profiling is a promising and efficient tool to determine intra-specific genetic variation among A. flavus populations from hospital environments. A. flavus isolates, either toxigenic or non-toxigenic, should be considered as potential threats for hospitalized patients due to their obvious role in the etiology of nosocomial aspergillosis.

The antibacterial activity of essential oils from caraway seeds together with its antioxidant properties were examined in seeds pretreated with γ-irradiation. The oil fraction components were not affected due to γ-irradiation as determined by gas chromatography/mass spectrometry. The antioxidant activities which were measured by β-carotene bleaching test and 2, 2-diphenylpicrylhydrazyl assay were sustained by γ-irradiation. The antibacterial activity of caraway essential oils was also retained after irradiation. These data may suggest that γ-irradiation sustains the active components responsible for the biochemical properties of the caraway oils. This could find additional application in food processing industries. PRACTICAL APPLICATION Caraway has been used as a popular aromatic herb and spice since antiquity and has been normally found in various parts of the world especially in Iran. Caraway seeds and its products such as essential oils are widely added to food and food products for flavoring and are also used in Iranian traditional medicine for treatment of different diseases. With regard to the contamination of the seeds by special microorganisms, their sanitization by a safe method such as γ-irradiation has a great application potential. The influences of irradiation on the active components of caraway seed, as a medicinal herb, responsible for the biochemical activities should not be ignored. Therefore, this study was designed to find out if γ-irradiation can alter the active components and biochemical properties of caraway seed preparations.

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 IC 50 values of AFB1 for hepatocytes differentiated from CD34 + and MSCs were within the same range (44.7–46.8 μM). The IC 50 calculated for non-differentiated MSCs and CD34 + cells was slightly lower (42.0–43.4 μM) 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 20 μM) for 24 h 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.

In this study, a new effective, pH and thermally stable glucose oxidase (GOX)–silver nanoparticles (AgNPs) bioconjugate was designed. AgNPs were synthesized based on the reduction of silver nitrate (AgNO 3 ) by sodium borohydride (NaBH 4 ) using two simple procedures. Periodic acid was used for oxidation of the GOX and emission of Lucifer yellow (LyCH) was monitored by spectrofluorometer for evaluation of the oxidation properties of the GOX. The oxidized GOX (Ox-GOX) was immobilized on AgNPs by its sugar moieties via 6-aminohexanoic acid (6AHA) as linker. A sample of the synthesized bioconjugate was loaded on 7.5% non-denaturing polyacrylamide gel electrophoresis (PAGE) to confirm its structural and physical stability. The results from enzymatic activity assay showed that the bioconjugate, GOX and Ox-GOX were similar in stability and activity in acidic and basic pH (optimum pH = 7.0–8.0). Based on the results from thermal stability assay, it was found that the activity of the bioconjugate was found to be higher at lower temperatures. The V max of the bioconjugate, GOX, and Ox-GOX was estimated as 28.6, 6.2, and 6 IU μg −1 enzyme and the K m was calculated as 2.7, 9, and 9.5 mM, respectively. It was found that the immobilization method improves the activity and stability of the GOX in different pH and temperatures. As a conclusion, the proposed method opens up the way to the development of a new bioconjugate with potential use in sensing, and many find potential applications in clinical diagnostics, medicine, and industries.

Background: Mesenchymal stem cells (MSCs), the non-hematopoietic progenitor cells found in various adult tissues are characterized by their ease of isolation and their rapid growth in vitro while maintaining their differentiation potential, allowing for extensive culture expansion to obtain large quantities suitable for therapeutic application. Silymarin has strong antioxidant and anti-inflammatory activities with positive effect on proliferation on some cell types. Objectives: The aim of this study was to find out the optimal condition of silymarin treatments on mesenchymal stem cell growth and multiplication. Methods: Human MSCs in third passage were divided into 12 groups treated by 50, 75, and 100 μg/ml of silymarin for 2, 7 and 14 days. Cell viability was assayed on day 2 using trypan blue exclusion test. The cell proliferation rate in presence of silymarin was determined using a day-response curve for each dose. Results: Viability was 89%, 93%, and 96% for cells treated with 50, 75, and 100µg/ml silymarin, respectively. Cell viability showed significant increase in all treated cells in comparison with the control group (83%). Based on the day- response curve, it was shown that the rate of cell proliferation in treated cells is significantly higher when exposed to silymarin for 2 - 7 days. However, from 7th day to 14th, silymarin exposure lowers hBMSCs proliferation rate compared to control group. Conclusion: Under optimal condition of silymarin exposure time, the rate of MSC proliferation can be stimulated.

BACKGROUND: The aim of this study was to investigate the effect of sodium selenite (SS) on reactive oxygen species (ROS) production, total antioxidant capacity (TAC) and glutathione peroxide (GPx) activity of cultured pre-antral follicles derived from vitrified and non-vitrified ovarian tissue. METHODS: Immature mouse ovaries were vitrified, and mechanically isolated pre-antral follicles from vitrified and non-vitrified samples were cultured in TCM 199 medium supplemented with different concentrations (0, 5 and 10 ng/ml) of SS. Follicular, oocyte and embryo development was assessed. In parallel, ROS, TAC and GPx levels were analyzed after 0, 12, 24, 48, 72 and 96 h of culture. RESULTS: Development rates of follicles, oocytes and embryos were significantly higher in SS-supplemented groups (P < 0.005). ROS production was increased, and TAC levels and GPx activities were decreased after 24 h of culture of pre-antral follicles in vitrified and non-vitrified groups, whereas in the presence of SS, ROS production was decreased and TAC levels and selenium-dependent GPx-specific activities were increased after 96 h of culture. Vitrified and non-vitrified samples responded in a similar manner. CONCLUSION: SS caused an increase in follicular TAC level and GPx activity and a decrease in ROS level, thus improving the in vitro development of follicles.

Irradiation is an important method of processing herbal drugs, while our understanding of the effects of gamma-irradiation on pharmacological properties of seed products such as caraway essential oils is however still very limited. In this study, caraway seeds were irradiated at dose levels of 0, 10 and 25kGy. After extracting the essential oils, the effects of fresh and gamma-irradiated caraway oils (100mg/kg b.w) on preventing septic-related oxidative liver injury induced by cecal ligation and puncture (CLP) model were investigated by measuring oxidative stress parameters in the liver. CLP operation caused a marked increase in myeloperoxidase (MPO) activity which was readily reversed in rats treated with fresh and irradiated caraway oils. Likewise, thiobarbituric acid reactive substances (TBARS) level in the liver was compensated in rats treated with the fresh and irradiated caraway oils. Moreover, liver GSH which was initially depleted due to CLP was recovered by essential oil treatments. The protective role of oils was further confirmed by showing that liver function tests (ALT/AST) as well as histopathological changes following CLP operation were recovered in rats treated with oils from either fresh or irradiated caraway seeds. These data may suggest that gamma-irradiation to caraway seeds at 10 and 25kGy has no influence on the antioxidative properties of caraway essential oils.

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.

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 (approximately 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.

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.

Aim To supporting growth and functional differentiation of adult stem cells into hepatocytes in a well-controlled manner, we performed differentiation of human bone marrow mesenchymal stem cells (hBMSCs) to hepatocytes-like cells on a constructed 3-dimensional (3D) nanofibrous biocompatible scaffold. Methods After characterization of the hBMSCs isolated from human bone marrow, the performance of the cells seeded and their proliferation on the scaffold was evaluated by scanning electron microscopy (SEM) and 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Different approaches such as immunocytochemistry, reverse transcriptase polymerase chain reaction (RT-PCR), and biochemical assays were used to estimate the ability of hBMSC-derived cells to express hepatocyte-specific markers. Results Scanning electron micrographs and MTT analysis revealed the cells were able to expand and remained biologically and metabolically active for 21 days. Immunocytochemical analysis of albumin and α-fetoprotein showing the accumulation of these markers in differentiated cells was confirmed by RT-PCR. Additional markers such as cytochrome P450 3A4, cytokeratin-18, and cytokeratin-19 detected by RT-PCR showed progressive expression during 3 weeks of differentiation on 3D scaffold. The hepatocyte-like cells displayed several characteristics of metabolic functions as judged by production of albumin, urea, transferrin, serum glutamic pyruvic transaminase (SGPT), and serum oxaloacetate aminotransferase (SGOT). Levels of above-mentioned markers, except SGOT in differentiated cells on scaffold, were found to be significantly greater than in the 2D culture system (pConclusion Overall data suggest that the engineered nanofibrous scaffold is a conductive matrix for functional hBMSC-derived hepatocyte-like cells and is promising for maintenance of hepatocytes suitable for implantation.

Aflatoxin B1 (AFB1) is a highly toxic and carcinogenic metabolite produced by Aspergillus species on food and agricultural commodities. Natural products may regulate the cellular effects of aflatoxins and evidence suggests that aromatic organic compounds of spices can control the production of aflatoxins. With a view to controlling aflatoxin production, the essential oils from Rosmarinus officinalis and Trachyspermum copticum L. were obtained by hydrodistillation. Antifungal activities of the oils were studied with special reference to the inhibition of Aspergillus parasiticus growth and aflatoxin production. Minimal inhibitory (MIC) and minimal fungicidal (MFC) concentrations of the oils were determined. T. copticum L. oil showed a stronger inhibitory effect than R. officinalis on the growth of A. parasiticus. Aflatoxin production was inhibited at 450 ppm of both oils with that of R. officinalis being stronger inhibitor. The oils were analyzed by GC and GC/MS. The major components of R. officinalis and T. copticum L. oils were Piperitone (23.65%), alpha-pinene (14.94%), Limonene (14.89%), 1,8-Cineole (7.43%) and Thymol (37.2%), P-Cymene (32.3%), gamma-Terpinene (27.3%) respectively. It is concluded that the essential oils could be safely used as preservative materials on some kinds of foods to protect them from toxigenic fungal infections.

To investigate the interaction between welding fumes and noise in causation of hearing impairment.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.The DPOAEs levels measured at different frequencies (1379-6299 Hz) were found to decrease significantly (P0.05) in rabbits exposed to 110 dB sound pressure level (SPL) broadband noise (8h/day, 12 days). In rabbits, exposed to carbon-steel welding fumes alone (157 mg/m(3)), the threshold shift was limited to the high frequencies (2759-6299 Hz), 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 (P0.05). 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.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.

Oxidative stress is a major factor in the pathogenesis of diabetic complications. We studied the effects of ginger powder on nephropathy induced by diabetes, and measured changes in plasma antioxidant capacity and lipid peroxidation. Wistar rats weighing 250 ± 20 g were treated with STZ 60 mg/kg. Rats were divided into 3 groups (n = 8) of non-diabetic, diabetic non-treated and diabetic treated with ginger powder. The diabetic treated with ginger group received ginger at 5% of their consumed food daily. After 8 weeks the rats were anaesthetized with 10% chloral hydrate. Blood samples were collected from the heart of each rat and kidneys were removed and kept in 10% formalin buffer. Plasma and red blood cells were separated. Plasma antioxidant capacity by the FRAP method and red blood cells malondialdehyde (MDA) as an indictor of lipids peroxidation, were measured. After sectioning and staining of renal samples, they were studied for focal cell proliferation and glomerular and tubular structural changes. The MDA levels in diabetic rats treated with ginger were significantly lower than in the other groups (P

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.

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

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(-1)) 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 B1 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(-1). 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.

The effects of Akacid(plus), a novel member of guanidine-based polymeric compounds recently introduced as a potent inhibitor of fungal growth and aflatoxin biosynthesis were studied on Aspergillus parasiticus by transmission electron microscopy (TEM). The toxigenic fungus was cultured on yeast extract-sucrose broth in presence of serial two-fold concentrations of Akacid(plus) (1.5-96 microL/50 mL medium) for 96 h at 28 degrees C with shaking. Mycelial samples exposed to fungistatic concentrations of compound (1.5-48 microL) were processed for TEM. Corresponding to the growth inhibition, TEM observations revealed morphological anomalies in fungal compartments. The results demonstrated that Akacid(plus) targets the plasma membrane of the hyphae by its breaking down at variable intervals with the formation of small membrane-bound vesicles inside the cytoplasm, while no obvious damage was observed on the cell wall. A marked depletion of cytoplasmic contents of hyphae accompanied with lysis and disruption of membranes of major organelles such as nuclei, mitochondria and endoplasmic reticulum indicates that in high fungistatic concentrations, Akacid(plus) passes not only through the cell wall but also through the plasma membrane and then interact with membranous structures of the cytoplasmic organelles. Ultrastructural changes of fungal compartments exposed to Akacid(plus) in relation to the fungal growth and aflatoxin biosynthesis are discussed.

Summary Objectives Listeria monocytogenes has gained increasing attention as a pathogen of public health importance owing to large numbers of food-borne outbreaks of listeriosis. Because of negative consumer perception of chemical preservatives, attention is shifting towards natural alternatives. Particular interest has been focused on the potential application of plant essential oils. The objective of the present study was to determine ultrastructural changes brought about by essential oils from two types of thyme, Thymus eriocalyx and Thymus x-porlock , on Listeria monocytogenes . Materials and methods Minimal inhibitory (MIC) and minimal bactericidal (MBC) concentrations and bactericidal kinetics of the oils were determined. Listeria monocytogenes were treated with essential oils from two thyme species and observed under a transmission electron microscope. Results The oils from the above plants were found to be strongly antimicrobial. Analysis of the oils by gas chromatography and gas chromatography/mass spectrometry lead to the identification of 18 and 19 components in T. eriocalyx and T. x-porlock oils, respectively. Listeria monocytogenes treated with essential oils from the two thyme species exhibited a thickened or disrupted cell wall with increased roughness and lack of cytoplasm. Conclusion The antilisterial effects of thyme oil are stronger than the action of electric shocks in combination with nisin reported in the literature. It is concluded that essential oils such as thyme oil, which inhibited the growth of L. monocytogenes at low concentrations, could be considered as preservative materials for some kinds of foods; they could find an application as additives to foodstuffs in storage to protect them from listerial contamination.

The antifungal effects of essential oils from Thymus eriocalyx and Thymus x-porlock were studied with special reference to the mechanism of inhibition of Aspergillus niger growth at ultrastructural level. Minimal inhibitory (MIC), minimal fungicidal (MFC) concentrations, and fungicidal kinetics of the oils were determined. Transmission electron microscopy (TEM) of A. niger exposed to MIC levels of the oils showed irreversible damage to cell wall, cell membrane and cellular organelles. The oils analyzed by GC and GC/MS led to identification of 18 and 19 components in T. eriocalyx and T. x-porlock oils respectively. The results are compared and discussed with the data in literature. It is concluded that the essential oils could be safely used as preservatives.

The methylotrophic yeast Pichia pastoris has become a highly popular expression host system for the recombinant production of a wide variety of proteins, such as antibody fragments. Camelids produce functional antibodies devoid of light chains and constant heavy-chain domain (CH1). The antigen binding fragments of such heavy chain antibodies are therefore comprised in one single domain, the so-called VH of the camelid heavy chain antibody (VHH). To test the feasibility of expressing VHHs in the yeast, which on account of their small size and antigen recognition properties would have a major impact on antibody engineering strategies, we constructed two VHH genes encoding the single-domain antibody fragments with specificity for a cancer associated mucin, MUC1. The recombinant strains of the yeast P. pastoris were developed which secrete single-domain antibody fragment to the culture supernatant as a biologically active protein. Supplementation of medium with sorbitol (in pre-induction phase) and casamino acid or EDTA (in induction phase) provided ideal condition of increasing the yield of VHH production compared to culture condition devoid of above recipe. The secreted protein was purified following a 80% ammonium sulfate precipitation step, followed by a affinity chromatography column. The specific activity in enzyme-linked immunosorbant assay (ELISA) of the purified yeast VHH was higher than that of a bacterial periplasmic counterpart. These results reaffirm that the yeast P. pastoris is a suitable host for high level and correctly folded production of VHH antibody fragments with potential in vivo diagnostic and therapeutic applications. This is the first report of expression of VHH in P. pastoris.

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, 1 ppm); group D, basal diet having ammonia treated aflatoxin contaminated (1 ppm) maize; group E, basal diet containing AFB (2 ppm) contaminated maize; group F, basal diet containing ammonia treated AFB (2 ppm) 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

The up-regulation of heat shock protein (HSP) expression has been proposed as a general biomarker of cellular protection against various environmental stresses and chemicals. The present study investigated the possibility of using HSP70 up-regulation as a biomarker of toxicant exposure in vitro. Cells of a rat hepatoma cell line (FGC4) were exposed to concentrations of 1,3-dichloroacetone, duroquinone, diquat dibromide, menadione, hydrogen peroxide, cadmium chloride (CdCl2) and sodium (meta)arsenite (NaAsO2) that elicited 20–50% cytotoxicity over a 24-hour period, and HSP70 levels were measured by ELISA. Up-regulation of HSP70 expression was demonstrated following treatment with menadione, CdCl2 and NaAsO2, but not with the other chemicals tested. A shorter exposure time (6 hours) and/or the use of non-toxic concentrations reduced the level of HSP70 up-regulation with menadione, CdCl2 and NaAsO2, but did not uncover any up-regulation with the other chemicals. Although the toxicity of the majority of the chemicals tested is believed to involve an oxidative stress component, the results of this study clearly demonstrate that up-regulation of HSP70 expression cannot be used as a general biomarker of toxicant exposure in vitro.

Cytochrome P450 1A1 (CYP1A1) is among the cytochrome P450 classes known to convert xenobiotics and endogenous compounds to toxic and/or carcinogenic metabolites. Suppression of CYP1A1 over expression by certain compounds is implicated in prevention of cancer caused by chemical carcinogens. Chemopreventive agents containing high levels of flavonoids and steroids-like compounds are known to suppress CYP1A1. This study was carried out for assessment of the genomic and proteomic effects of caraway (Carum carvi) extracts containing high levels of both flavonoids and steroid-like substances on ethoxy resorufin dealkylation (EROD) activity and CYP1A1 at mRNA levels. Rat hepatoma cells co-treated with a CYP1A1 inducer i.e. TCDD (2, 3, 7, 8-tetrachlorodibenzo-p-dioxin) and different preparations of caraway extracts at concentrations of 0, 0.13, 1.3, and 13 microM in culture medium. After incubation (37 degrees C and 7% CO2 for 20 h), changes in EROD specific activity recorded and compared in cells under different treatments. The results show that caraway seed extract prepared in three different organic solvents suppressed the enzyme activity in hepatoma cells in a dose-dependent manner. The extracts added above 0.13 microM could significantly inhibit EROD activity and higher levels of each extract (1.3 and 13 microM) caused approximately 10-fold suppression in the enzyme activity. Accordingly, data obtained from the RT-PCR (TaqMan) clearly showed the suppressive effects of plant extract on CYP1A1-related mRNA expression. These data clearly show that substances in caraway seeds extractable in organic solvents can potentially reverse the TCDD-dependent induction in cytochrome P450 1A1.

To develop an indirect enzyme-linked immunosorbent assay (ELISA) for measuring class-pi glutathione S-transferase (GST) in plasma, and tissue biopsies obtained from upper gastrointestinal cancer (UGI Ca) patients.GST activity and GST-pi concentration were detected in normal human squamous esophageal epithelium, normal gastric cardia and their corresponding malignant tumor biopsies.Plasma GST was significantly higher (p0.05) in UGI Ca patients as compared to those obtained from normal individuals. Plasma GST-pi concentration in normal subjects was 6.6 +/- 1.9 ng/mg protein, whereas it was higher in UGI Ca patients (esophageal, 10.0 +/- 1.8; gastric, 10.7 +/- 1.7 ng/mL, por= 0.01). GST and GST-pi levels were higher in surgically resected tumor biopsies as compared to normal tissues adjacent to a tumor (p0.05). GST-pi was also elevated in malignant esophagus and gastric biopsies taken at endoscopy as compared to normal and normal-appearing esophageal tissues (p0.05). Total GST was also increased significantly in gastric malignant tissues although its activity was within the same range in normal and normal-appearing tissues. A significant correlation between plasma GST-pi with that in malignant tissues was observed. Total GST and GST-pi were also correlated in different malignant tissues.Biopsies obtained at endoscopy can be used satisfactorily to measure tumor markers.

Previous studies from this laboratory indicated that glutathione (GSH) conjugate formation with acetaminophen (APAP) is remarkably induced in liver of weanling rats in response to a single overdose of the drug administered intraperitoneally (ip). Increased APAP-GSH conjugation has been attributed to inducible glutathione S-transferases (GSTs) in dividing hepatocytes. In order to verify this finding, an in vitro reconstitution assay containing liver microsomes (source of cytochrome P-450) and cytosolic fractions (source of GST) from livers and kidneys of adult and weanling rats has been established. In vitro incubation of the reaction mixture was followed by solvent extraction, enzymatic digestion and HPLC analysis of the conjugate. Under controlled conditions, in vitro, the rate of APAP-GSH conjugation reflected the GST activity of cytosolic sample added to incubation system. The activity of cytosolic GST in catalyzing this reaction was measured using cytosols prepared from various tissue sources, particularly from animals pretreated with dietary butylated hydroxylanisole (BHA). The extent of APAP-GSH conjugate formation mediated by cytosols varied in this order: BHA-treated adult liver>BHA-treated weanling liver>control adult liver>control weanling liver>BHA-adult kidney>control adult kidney>BHA weanling kidney>control weanling kidney. In contrast to findings obtained from in vivo experiments, the rate of GST-dependent APAP conjugate formation with GSH in vitro is not induced in the presence of exogenous drug.

The relationship between the activities of 3 cytosolic enzymes with aflatoxin biosynthesis in Aspergillus parasiticus cultured under different conditions has been investigated in order to find out the role of each enzyme in aflatoxin biosynthesis. Basically the activity of isocitrate dehydrogenase (IDH) was higher in non-toxigenic strains as compared to its counterpart toxigenic fungi (p 90% inhibition) when the concentration of neem extract was adjusted to 50% (v/v). No significant changes in FAS and IDH activities were observed when aflatoxin synthesis was under restraints by neem (Azadirachta indica) leaf extract. During a certain period of time of culture growth, when aflatoxin production reached to its maximum level, the activity of FAS was slightly induced in the toxigenic strains fed with a low concentration (1.56% v/v) of the neem leaf extract. At the time (96 h) when aflatoxin concentration reached to its maximum levels, the activity of GST in the toxigenic fungi was significantly higher (i.e., 7–11 folds) than that of non-toxigenic strains. The difference was highest in mycelial samples collected after 120 h. However unlike FAS and IDH, GST was readily inhibited (∼67%) in mycelia fed with 1.56% v/v of the neem extract. The inhibition reached to maximum of 80% in samples exposed to 6.25–12.5% of the extract. These results further substantiate previous finding that there is a positive correlation between GST activity and aflatoxin production in fungi.

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.

International audience; Soil isolates of Aspergillus section Flavi from Mazandaran and Semnan provinces with totally different climatic conditions in Iran were examined for aflatoxins (AFs; B and G types), cyclopiazonic acid (CPA) and sclerotia production. A total of 66 Aspergillus flavus group strains were identified from three species viz. Aspergillus flavus, Aspergillus parasiticus and Aspergillus nomius in both locations. A. flavus (87.9%) was found to be the prominent species followed by A. nomius (9.1%) and A. parasiticus (3.0%). Only 27.5% of A. flavus isolates were aflatoxigenic (B(1) or B(1) and B(2)), out of which approximately 75% were capable to producing CPA. All the A. parasiticus and A. nomius isolates produced AFs of both B (B(1) and B(2)) and G (G(1) and G(2)) types, but did not produce CPA. Sclerotia production was observed in only 4 isolates of A. flavus among all 66 isolates from three identified species. A. flavus isolates were classified into various chemotypes based on the ability to produce aflatoxins and CPA. In this study, a new naturally occurring toxigenic A. flavus chemotype comprising of two strains capable of producing more AFB(2) than AFB(1) has been identified. A relatively larger proportion of aflatoxigenic A. flavus strains were isolated from corn field soils of Mazandaran province which indicate a possible relationship between high levels of relative humidity and the incidence of aflatoxin-producing fungi. The importance of incidence of Aspergillus section Flavi in corn field soils regard to their mycotoxin production profiles and crop contamination with special reference to climatic conditions is discussed.

The rate of glutathione (GSH) conjugate formation to acetaminophen (APAP) in livers of weanling and adult rats treated with a single i.p. dose of APAP was compared. HPLC analysis of cytosolic fractions revealed that rate of conjugation in weanling rat is 24-times greater than that of adults. Increased rate of GSH conjugation was independent of of the age-related difference observed in liver GSH content. The normal level of liver GSH in weanling rat was 57% of adult level. APAP treatment depleted GSH more significantly in weanling rats as compared to that in adults. N-acetylcystein (NAC) alone had little influence on liver GSH levels. However it was successful in reducing GSH depletion in tissues of growing rats. A 32% repletion in hepatic GSH level in NAC-treated weanling rats was associated with a further 13-fold increase in the rate of GSH conjugate formation. These data together with histopathological results, clearly showed that the inducible GSH system in weanling rat liver act as a safe guard against APAP toxicity. A surge in the rate of APAP-GSH conjugation in growing liver may function in compensation of other detoxification pathways which are saturated more readily at this age.

Noise induced hearin gloss (NIHL) is one of the most important occupational disease world wide. NIHL has been found potentiate by simultaneous carbon monoxide (CO) exposure. Free radicals have been implicated in cochlear damage resulted from the exposure to noise and due to the CO hypoxia. This study examined whether α-tocopherol administration , as a free radical scavenger, causes the attenuation of auditory brainstem response (ABR) threshold shifts resulting from noise exposure and noise plus CO exposure. Forty-two rabbits were divided in to seven groups including control, noise + saline, noise + CO + saline, noise + α-tocopherol, noise + CO + α-tocopherol , CO + α-tocopherol and α-tocopherol alone. ABR was assessed before exposure, 1 hand 14 days post exposure. The administration of 50 mg/Kg of α-tocopherol prior, following and post exposure to noise or noise plus CO recovered permanent ABR threshold shift at 1 and KHz almost to the baseline and provided significant attenuation in permanent ABR threshold shift at 4 and 8 KHz in subject swhich were exposed to noise but it did not block the potentiating of threshold elevation by CO exposure (extra threshold loss by combined exposure) at 4 and 8 KHz. α Tocopherol provides protective effect against the hearing loss resulting from noise exposure and simultaneous exposure to noise plus CO.

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.

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.

Background: Nosocomial fungal infections could arise from independent exposure to airborne spores of filamentous fungi existing in the hospital environment. Objectives: The present study aimed to determine the mycoflora of indoor and outdoor environments of five major hospitals in Khorramabad, Iran. Materials and Methods: Sampling of air was done from indoor and outdoor environments of wards, surroundings and green space of hospitals by settle plate method. To obtain the sample from surfaces, pre-moistened swabs with cotton-tipped sticks were applied on different surfaces (floor, the walls, windows, beds, trolleys, laryngoscope and angiography devices). Culture plates of air and surfaces on Potato Dextrose Agar (PDA) and Malt Extract Agar (MEA) were incubated in the dark at 28 C and examined daily for fungal colonies for two to three weeks. Fungal isolates were identified by a combination of their macroscopic and microscopic criteria after purification on isolation culture media. Results: A total of 707 fungal colonies including, Penicillium (29.14%), Cladosporium (24.04%), Aspergillus (20.65%), Fusarium (9.05%), Alternaria (3.96%), Rhodotorula (1.69%), Cryptococcus neoformans (0.7%) and other fungi (10.77%) were isolated. All the examined high-risk parts of the hospitals were found to be contaminated by various fungi. Conclusions: Aspergillus was the most prominent genus in Intensive Care Unit (ICU) and surgery, Cladosporium in Critical Care Unit (CCU), emergency and thalassemia, and Penicillium in orthopedic, emergency and neonatal sections. Among pathogenic yeasts, C. neoformans was isolated from ICU, surgery and orthopedic sections. The dimorphic fungal pathogen, Sporothrix schenckii, was reported from CCU. The isolated fungi specially the genera Aspergillus and Penicillium are potential threats for immunocompromised patients in the hospitals.

We all are exposed through air, food, drinks and skin contacts to harmful compounds throughout the period of our lifetime, including, a variety of pharmaceuticals and food-de‐ rived carcinogen metabolite (e.g. N-acetoxy-PhIP), [52], plant toxins (such as glycoalkaloids in nightshades1, cyanogenic glucosides 2, or pyrrolizidine alkaloids in some herbs and herbal teas), xenobiotics3 producing during early human pregnancy, fungal and bacterial toxins such as aflatoxins4; and cyanotoxin5; as well as free radicals and hydroperoxides. Many of these compounds are lipophilic and the organism can get rid of them only through metabolism.

In this study, the ability of neonatal rat liver to metabolise [3H]aflatoxin B1 (AFB1) was compared to that of the adult animal. In order to make this comparison, neonatal and young adult rats were killed 2, 6, 12 and 24 h after injection with a single i.p. dose of AFB1. The rate of AFB1 adduct formation to nuclear DNA and protein was measured in hepatic and pulmonary tissues. The results demonstrated that AFB1 was epoxidized more rapidly by the adult's liver and lungs 2 h after the toxin administration, compared with those of the neonatal's (adult 30 pmol and neonatal 12 pmol AFB1 bound/mg DNA). However, these differences were more pronounced in hepatic than in pulmonary tissues. The same differences between AFB1-DNA adducts were also observed at different time points. These changes are certainly related to the level of hepatic cytochrome P-450. The delayed cytochrome P-450-dependent AFB1 activation in neonatal's liver provides time enough for de-epoxidation of slowly generated epoxide. The rate of AFB1-epoxide formation at this age was consistent with the activity of phase II metabolism of AFB1 (glutathione conjugation). In addition, the hydrolysis of AFB1-DNA adducts at a relatively higher rate by neonatal's liver may also contribute to the quick removal of the adducts. In spite of the aforementioned evidence which shows the capability of neonatal liver to handle AFB1, the fate of large amounts of free (non-metabolised) AFB1 deposited in neonatal's liver is not well understood.

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.