The effect of deoxynivalenol on the secretion activity,proliferation and apoptosis of porcine ovarian granulosa cells in vitro

The aim of this in vitro study was to examine the secretion activity, markers of proliferation and apoptosis in porcine ovarian granulosa cells (GCs) after deoxynivalenol (DON) addition. Ovarian granulosa cells were incubated with DON for 24h: 10, 100 and 1000 ng/mL, while the control group received no DON. The secretion of insulin-like growth factor I (IGF–I) and progesterone was determined by radioimmunoassay (RIA) and expression of cyclin B1, PCNA and caspase-3 by immunocytochemistry. IGF–I release by GCs was inhibited by DON, while progesterone release and the expression of cyclin B1 was stimulated by DON (at 1000 ng/mL but not at 10 and 100 ng/mL). PCNA expression was stimulated by DON (at 100 and 1000 ng/mL but not at 10 ng/mL). Caspase-3 expression was not influenced by DON treatment (at all doses). In conclusion, our results indicate, (1) a direct effect of DON on secretion of growth factor IGF-I and steroid hormone progesterone, (2) expression of markers of proliferation (cyclin B1 and PCNA) but not on the (3) expression of marker of apoptosis (caspase-3) in porcine ovarian granulosa cells. This in vitro study suggests the dose-dependent association of DON on porcine ovarian functions.     

The effect of deoxynivalenol on the secretion activity, proliferation and apoptosis of porcine ovarian granulosa cells in vitro

The aim of this in vitro study was to examine the secretion activity, markers of proliferation and apoptosis in porcine ovarian granulosa cells (GCs) after deoxynivalenol (DON) addition. Ovarian granulosa cells were incubated with DON for 24h: 10, 100 and 1000 ng/mL, while the control group received no DON. The secretion of insulin-like growth factor I (IGF-I) and progesterone was determined by radioimmunoassay (RIA) and expression of cyclin B1, PCNA and caspase-3 by immunocytochemistry. IGF-I release by GCs was inhibited by DON, while progesterone release and the expression of cyclin B1 was stimulated by DON (at 1000 ng/mL but not at 10 and 100 ng/mL). PCNA expression was stimulated by DON (at 100 and 1000 ng/mL but not at 10 ng/mL). Caspase-3 expression was not influenced by DON treatment (at all doses). In conclusion, our results indicate, (1) a direct effect of DON on secretion of growth factor IGF-I and steroid hormone progesterone, (2) expression of markers of proliferation (cyclin B1 and PCNA) but not on the (3) expression of marker of apoptosis (caspase-3) in porcine ovarian granulosa cells. This in vitro study suggests the dose-dependent association of DON on porcine ovarian functions.     

Assessment of a potential preventive ability of amygdalin in mycotoxin-induced ovarian toxicity

The possible effects of a natural substance amygdalin and its combination with the mycotoxin deoxynivalenol (DON) on the steroid hormone secretion (progesterone and 17-β-estradiol) by porcine ovarian granulosa cells (GCs) were examined in this in vitro study. Ovarian GCs were incubated without (control group) and with amygdalin (1, 10, 100, 1,000 and 10,000 μg mL¹), or its combination with DON (1 μg mL¹) for 24 h. The release of steroid hormones was determined by ELISA. The progesterone secretion by porcine ovarian GCs was not affected by amygdalin in comparison to the control. However, the highest amygdalin dose (10,000 μg mL¹) caused a significant stimulation of the 17-β-estradiol release. A combination of amygdalin with DON significantly (P < 0.05) increased the progesterone release at all concentrations. Similarly, a stimulatory effect of amygdalin co-administered with DON was detected with respect to the 17-β-estradiol secretion at the highest dose (10,000 μg mL¹) of amygdalin and 1 μg mL¹ of DON. Noticeable differences between the effects of amygdalin alone and its combination with DON on the progesterone release were detected. In contrast, no differences between the stimulatory effects of amygdalin and its combination with DON on the 17-β-estradiol synthesis by porcine GCs were observed. Findings from this in vitro study did not confirm the expected protective effect of amygdalin on mycotoxin induced reprotoxicity. Our results indicate that the stimulatory effect of amygdalin combined with DON on the progesterone release was clearly caused by the DON addition, not by the presence amygdalin per se. On the other hand, the stimulation of 17-β-estradiol production was solely caused by the presence of amygdalin addition. These findings suggest a possible involvement of both natural substances into the processes of steroidogenesis and appear to be endocrine modulators of porcine ovaries.     

Dietary bioflavonoid quercetin modulates porcine ovarian granulosa cell functions in vitro

Quercetin is a dietary bioflavonoid used widely as a food supplement and is generally recognized as safe. The aim of this in vitro study was to examine the steroid hormone (progesterone and 17- β estradiol) release, proliferation (PCNA and cyclin B1) and apoptosis (caspase 3 and p53) of porcine ovarian granulosa cells after the addition of quercetin at concentrations 0.01, 0.1, 1, 10 and 100?μmol?L^?1. Progesterone release was stimulated at the concentration 10?μmol?L^?1. Quercetin neither had any impact on 17-β estradiol secretion nor on the presence of PCNA. However, a significant enhancement of the occurrence of cyclin B1 was noted except for the lowest concentration 0.01?μmol?L^?1. Quercetin did not have any influence on the number of granulosa cells containing caspase 3, but at the concentration 10?μmol?L^?1 it inhibited p53 occurrence. Results confirm the safety of quercetin in porcine ovarian granulosa cell model and further suggest its possible concentration-dependent influence on ovarian functions through pathway that may involve progesterone, cyclin B1 and p53.     

The effect of bee pollen on secretion activity, markers of proliferation and apoptosis of porcine ovarian granulosa cells in vitro

The general objective of this in vitro study was to examine the effect of bee pollen on the release of insulin-like growth factor I (IGF-I) and steroid hormone progesterone, and expression of markers of proliferation (PCNA) and apoptosis (caspase-3) in porcine ovarian granulosa cells. Concentrations of IGF-I and progesterone were determined by RIA method and expression of PCNA and caspase-3 by immunocytochemistry. Bee pollen addition at the dose of 10 ng/mL significantly (P<0.05) inhibited IGF-I release by porcine ovarian granulosa cells. This growth factor was not influenced by 100 and 1000 ng/mL doses of bee pollen. Progesterone release by cells was not influenced by bee pollen addition at the doses of 10, 100 and 1000 ng/mL as used in our study. Similarly expression of PCNA and caspase-3 was not affected by bee pollen addition. The present study shows dose-dependent regulation of IGF-I by experimental bee pollen addition in vitro. Progesterone release, expression of PCNA and caspase-3 in porcine ovarian granulosa cells was not induced by pollen. Our results contribute to new insights regarding the possible effect of bee pollen on IGF-I release, which is important for regulation of porcine ovarian functions.     

Deactivation of mycotoxins. I. An in vitro study of zearalenone adsorption on new polymeric adsorbents

Abstract

This study describes the elimination of zearalenone concentrations in vitro using two new polymeric forms of cross‐linked polyvinylpyrrolidone (cryogels of cross‐linked PVP). Adsorption of zearalenone was studied under isothermal conditions and simulating pH of intestinal environment. A Freundlich isotherm was used to describe the adsorption data obtained. The results showed significant decrease of zearalenone concentrations, ranging from 33.5–66.2 % per 25 mg of polymer. Adsorption capacity (k) was estimated to be higher than that of previously tested adsorbents, including crospovidone. The data indicate the need to investigate structure peculiarities in order to improve mycotoxin deactivation procedures using PVP derivatives.     

Transepithelial transport and cellular accumulation of steroid hormones and polychlorobiphenyl in porcine kidney cells expressed with human P-glycoprotein

Endocrine disrupters such as sex hormone-like chemicals and the non-physiological ligands for aryl hydrocarbon receptor (AhR) exert many adverse biological effects. The ligands for AhR disturb gene expression downstream of the gene induced by estrogen receptor at a very low concentration. Thus, transepithelial transport and cellular accumulation of cortisol (COR) and estrogen as congeners of sex hormone-like chemicals, and 3,3^′,4,4^′-tetrachlorobiphenyl (TeCB) as one of the ligands for AhR were examined in a monolayer of porcine kidney cells transfected with human P-glycoprotein (LLC-COL). The net basal-to-apical transport of COR increased in LLC-COL compared to that in the wild type cells (LLC-PK1) the same as in vinblastine, whereas the net transport of estradiol (EST) was not detected in either cell group. Though the diffusion transports of EST for both directions, basal-to-apical and apical-to-basal, were higher than that of COR, cellular accumulation of EST was higher than that of COR. Transepithelial transport of TeCB was very low and the net basal-to-apical transport was not detected, while it was highly accumulated in the epithelial cells. The accumulation was slightly higher in LLC-COL than in LLC-PK1 at high dose.     

Low dose exposure of patulin and protective effect of epicatechin on blood cells in vitro

In the present study, we aimed to assess antioxidant status in erythrocytes in vitro after patulin (PAT) and epicatechin exposure by measuring antioxidant enzymes (superoxide-dismutase – SOD, glutathione peroxidase – GPx and catalase – CAT) and parameters associated with oxidative stress (malondialdehyde – MDA and ROS). We also investigated the effect of PAT on viability and count of lymphocytes and lymphocyte subpopulations in rabbit blood in vitro. Whole blood of rabbits was used for analysis of antioxidant changes in rabbit erythrocytes after epicatechin and PAT treatment (separately or in combination, at concentrations of 0.2; 2; 20; 200?µg mL^–1 of epicatechin and 0.5; 5; 10?µg mL^–1 of PAT). Whole blood of rabbits was also used for analysis of count and viability of lymphocytes after PAT treatment at concentrations of 10; 25 and 50?µg mL^–1. Results from our experiment confirmed the ability of epicatechin to protect cells against oxidative stress and lipoperoxidation. Our findings indicate that mycotoxin PAT in low concentrations did not affect the activity of antioxidant enzymes in erythrocytes of rabbits significantly. Only slight non-significant changes in lymphocytes count after treatment with low doses of PAT in rabbit blood were observed.     

The influence of lindane and lindane metabolites on microsomal and mitochondrial ATPase in vitro

In vitro investigations of the influence of lindane and its metabolites were performed on microsomal and mitochondrial ATPases from liver, kidney and brain of rat and mouse. The microsomal Na+-K+-ATPases in rat liver were inhibited by the tested substances. An increase of activity was observed only with 2.5 X 10(-5) M gamma-HCH. Effects on the microsomal Na+-K+-ATPase from kidney and brain of rat were also indicated. The mitochondrial enzyme in rat liver was stimulated by all the compounds tested at concentrations of 10(-4) M - 10(-2) M. The effects on mitochondrial enzymes from kidney and brain varied in dependence on the tested substances. In the microsomes and mitochondria of mouse an influence on the Na+-K+-ATPases similar to the effects on the preparations from organs of rat was evident.     

Doramectin induced cytotoxic and genotoxic effects on bovine peripheral lymphocytes and cumulus cells in vitro

The effect of doramectin (DOR) was tested on two experimental somatic bovine cells in vitro: peripheral lymphocytes (PL) and cumulus cells (CC). The cytotoxicity and genotoxicity of DOR were assessed using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, single cell gel electrophoresis assay (SCGE) and cytokinesis-block micronucleus cytome (CBMN Cyt) assay. Both cells were treated with three concentrations of DOR (20, 40, 60?ng?mL^–1) for 24?h. The results obtained from PL demonstrated that DOR was able to induce cytotoxic effect and DNA damage with all concentrations tested. Additionally, DOR increased micronuclei (MNi) frequency and nuclear buds (NBuds) with 20, 40, 60?ng?mL^–1, and nucleoplasmic bridges (NPBs) only with 40?ng?mL^–1. On the other hand, the three concentrations of DOR were not able to induce cytotoxic effect and DNA damage using SCGE in the bovine CC. Nevertheless, the two higher concentrations of DOR (20, 40?µg mL^–1) significantly increased the frequency of micronucleus formation in bovine CC. These results represent the first experimental evidence of genotoxic and cytotoxic effects exerted by DOR on bovine PL and CC.     

甲基叔丁基醚(MTBE)对环境的污染及其对我国汽油生产的影响

本文简要介绍了汽油添加剂MTBE对环境的污染及减少MTBE污染机理研究的进展 ,同时介绍了对我国汽油生产的影响     

The in vitro effect of selected essential oils on the growth and mycotoxin production of Aspergillus species

The aim of the present study was to assess the antifungal and anti-toxinogenic activity of 15 essential oils (EOs) against three fungi of the genus Aspergillus (A. parasiticus KMi-227-LR, A. parasiticus KMi-220-LR and A. flavus KMi-202-LR). The minimum inhibitory doses (MIDs) of the tested essential oils and their antifungal activity were determined using the micro-atmosphere method. The original commercial essential oil samples of Jasminum officinale L., Thymus vulgaris L., Syzygium aromaticum (L.) Merrill &; Perry, Rosmarinus officinalis L., Ocimum basilicum L., Eucalyptus globulus Labill., Salvia officinalis L., Citrus limon (L.) Burm, Origanum vulgare L., Lavandula angustifolia Mill., Carum carvi L., Citrus sinensis (L.) Osbeck., Zingiber officinalis Rosc., Mentha piperita L. and Cinnamomum zeylanicum Nees. (C. verum J.S.Presl.) were produced in Slovakia (Calendula a.s., Nová ?ubovňa, Slovakia). All essential oils exhibited activity against all tested strains of fungi. After 14 days of incubation, A. flavus (KMi-202-LR) showed the highest susceptibility with a growth inhibition percentage (GIP) of 18.70% to C. limon and 5.92% to C. sinensis, while A. parasiticus (KMi-220-LR) exhibited a GIP of 20.56% to J. officinale. The minimum inhibitory doses (MIDs) of EOs with the most significant activity were recorded. The best antifungal activity, using the micro-atmosphere method was found in S. aromaticum with an MID of 62.5 μL L^?1 air, T. vulgaris (MID of 62.5 μL L^?1 air) and O. vulgare (MID of 31.5 μL L^?1 air) against all tested strains. Mycotoxin production of the tested strains was evaluated by the thin layer chromatography (TLC) method. Mycotoxin production of AFB₁ and AFG₁ was inhibited following all treatments with C. carvi, R. officinale and S. officinale, Eucalyptus globulus L. and O. basilicum L. Essential oils exhibited a potential inhibition activity against toxic fungi, although, these affected only the production of AFB₁.     

水源地生态工程对消毒副产物生成的影响

水源地生态工程可改善饮用水水源水质,但其中的水生生物代谢物可能是消毒副产物(DBPs)前体物的来源。本文构建现场实验装置探究了水源地生态工程对原水水质的影响及原水中主要消毒副产物前体物的来源,考察了氯投量、温度以及pH对香蒲根分泌物、菰根分泌物和鲢鱼排泄物氯化后消毒副产物生成的影响。结果表明,实验装置对NH₄⁺-N、TN和TP的平均总去除率分别为74.93%、53.98%和73.02%,总DOC沿程增加。溶解性有机物(DOM)中分子质量分布在<500 Da的DOC含量总体上呈沿程减少的趋势,>3 000 Da的DOC沿程有所增加。总三卤甲烷生成势(TTHMFP)和总卤乙酸生成势(THAAFP)沿程呈现增加的趋势,分子质量<3 000 Da的有机物中TTHMFP和THAAFP沿程有所下降,分子质量>3 000 Da的TTHMFP和THAAFP呈沿程增加的趋势。考察了装置中3种水生生物代谢物经氯化后得到的二氯乙酸(DCAA)、三氯乙酸(TCAA)、三氯甲烷(TCM)和二氯乙腈(DCAN)4种消毒副产物生成势,均随着氯投量和温度的增加而升高,主要消毒副产物为TCAA。酸性条件有利于抑制DCAA和TCM的生成,碱性条件有利于抑制TCAA的生成,DCAN的生成量总体上随着pH的升高呈现先上升后下降的趋势。     

The effects of benomyl and its breakdown products carbendazim and butyl isocyanate on the structure and function of tracheal ciliated cells

Abstract

The effects of the fungicide benomyl and its breakdown products, carbendazim and butyl isocyanate, were examined on canine tracheal epithelial tissue in primary culture. Changes in ciliary frequencies were monitored with an optical spectrum analysis system. Serial dilutions of the test compounds were prepared in 100% corn oil and applied to the cell cultures for intervals up to 6 hours and frequencies measured at intervals of 15 minutes to 1 hour. Benomyl and butyl isocyanate caused concentration‐dependent decreases in ciliary beat frequency. Benomyl at 300 μg/ml (3 mM) caused ciliostasis within 75 minutes of exposure. Butyl isocyanate at a molar concentration three times lower than benomyl (1 mM) caused a similar response, although within 30 minutes. The IBC₅₀ for benomyl was 0.75 mM, while for butyl isocyanate it was 0.52 mM. Carbendazim caused a moderate decrease in frequency over a 6 hour exposure period. Benomyl caused moderate to severe swelling of the mitochondria of ciliated epithelial cells with other cell organelles appearing normal. Butyl isocyanate did not cause any noticeable effect on cell ultrastructure and the apparently low rate of penetration of carbendazim into cells made it impossible to obtain an effect which justified ultrastructural analysis. It appears, at least for benomyl and butyl isocyanate, that while the physiological effect of these two compounds (inhibition of ciliary beat) is the same, the sites of action in the cell may be different.     

Influence of herbicide glyphosate on growth and aflatoxin B1 production by Aspergillus section Flavi strains isolated from soil on in vitro assay

The effect of six glyphosate concentrations on growth rate and aflatoxin B₁ (AFB₁) production by Aspergillus section Flavi strains under different water activity (a_W) on maize-based medium was investigated. In general, the lag phase decreased as glyphosate concentration increased and all the strains showed the same behavior at the different conditions tested. The glyphosate increased significantly the growth of all Aspergillus section Flavi strains in different percentages with respect to control depending on pesticide concentration. At 5.0 and 10 mM this fact was more evident; however significant differences between both concentrations were not observed in most strains. Aflatoxin B₁ production did not show noticeable differences among different pesticide concentrations assayed at all a_W in both strains. This study has shown that these Aspergillus flavus and A. parasiticus strains are able to grow effectively and produce aflatoxins in high nutrient status media over a range of glyphosate concentrations under different water activity conditions.     

有机砷对固态厌氧消化产甲烷的影响和动力学研究

通过高温(55℃)固态厌氧消化研究,发现添加洛克沙胂(ROX)会抑制固态厌氧消化产甲烷,ROX添加水平为0.5、1.0、1.5mmol/kg时,初始每克挥发性固体(VS)累积产甲烷量依次下降19.19%、94.04%、96.56%;添加3-氨基-4-羟基苯砷酸(HAPA)加快了系统前期产甲烷,最终累积产甲烷量与空白对照组相近,抑制效应不明显。一级动力学模型符合固态厌氧消化系统产甲烷动力学,其基本方程为G=G∞×(1-exp(-kt)),其中:G为试验检测得到的累积产甲烷量,mL;G∞为理想状态下基质完全降解时的累积产甲烷量,mL;k为产气一级反应速率常数,d-1;t为反应时间,d。一级动力学模型预测值与实测值的F检验以及P和r2说明,产甲烷动力学方程与实测数据拟合相关度很高,产甲烷动力学方程的回归是极显著的。     

The influence of long-term fertilization on cadmium (Cd) accumulation in soil and its uptake by crops

Continuous application of organic and inorganic fertilizers can affect soil and food quality with respect to heavy metal concentrations. The risk of cadmium (Cd) contamination in a long-term (over 20 years) experimental field in North China with an annual crop rotation of winter wheat and summer maize was investigated. The long-term experiment had a complete randomized block design with seven fertilizer treatments and four replications. The seven fertilizer treatments were (1) organic compost (OM), (2) half organic compost plus half chemical fertilizer (OM?+?NPK), (3) NPK fertilizer (NPK), (4–6) chemical fertilizers without one of the major nutrients (NP, PK, and NK), and (7) an unamended control (CK). Soil samples from 0 to 20 cm were collected in 1989, 1999, and 2009 to characterize Cd and other soil properties. During the past 20 years, various extents of Cd accumulation were observed in the soil, and the accumulation was mainly affected by atmospheric dry and wet deposition and fertilization. In 2009, the average Cd concentration in the soil was 148?±?15 μg kg^?1 and decreased in the order of NPK?≈?OM?+?NKP?≈?PK?>?NP?≈?NK?>?OM?≈?CK. Sequential extraction of Cd showed that the acid-soluble fraction (F1, 32?±?7 %) and the residual fraction (F4, 31?±?5 %) were the dominant fractions of Cd in the soil, followed by the reducible fraction (F2, 22?±?5 %) and oxidizable fraction (F3, 15?±?6 %). The acid-soluble Cd fraction in the soil and Cd accumulation in the crops increased with soil plant available K. Fraction F3 was increased by soil organic C (SOC) and crop yields, but SOC reduced the uptake of soil Cd by crops. The long-term P fertilization resulted in more Cd buildup in the soil than other treatments, but the uptake of Cd by crops was inhibited by the precipitation of Cd with phosphate in the soil. Although soil Cd was slightly increased over the 20 years of intensive crop production, both soil and grain/kernel Cd concentrations were still below the national standards for environmental and food safety.     

The effect of emissions of fertilizer production on the environment contamination by cadmium and arsenic in southern Brazil

The distribution of cadmium and arsenic in rainwater, surface soil and groundwater in a zone of influence of emissions of a phosphate fertilizer factory has been investigated. The analysis of rainwater along a wind line shows that cadmium in the soluble form precipitates close to emission source, whereas arsenic migrates further and also in the soluble form precipitates at a greater distance. The pH controls the distribution of cadmium and arsenic in rain and groundwater. Concentration of soluble cadmium along the impact line increases with reduction of pH and arsenic, contrary to the increase of pH. Concentrations of cadmium and arsenic in surface soil correlate well among themselves near to emission point. In a zone of confluence of emissions with sea air at a distance from the factory there is sedimentation of arsenic on a geochemical barrier and formation of a separate anomaly in soil.     

The influence of pH and organic matter content in paddy soil on heavy metal availability and their uptake by rice plants

The experiments were done to investigate the effect of soil pH and organic matter content on EDTA-extractable heavy metal contents in soils and heavy metal concentrations in rice straw and grains. EDTA-extractable Cr contents in soils and concentrations in rice tissues were negatively correlated with soil pH, but positively correlated with organic matter content. The combination of soil pH and organic matter content would produce the more precise regression models for estimation of EDTA-Cu, Pb and Zn contents in soils, demonstrating the distinct effect of the two factors on the availability of these heavy metals in soils. Soil pH greatly affected heavy metal concentrations in rice plants. Furthermore, inclusion of other soil properties in the stepwise regression analysis improved the regression models for predicting straw Fe and grain Zn concentrations, indicating that other soil properties should be taken into consideration for precise predicting of heavy metal concentrations in rice plants.     

脱氮副球菌YF1微生物燃料电池生物阴极脱氮和产电

以脱氮副球菌YF1构建纯种生物阴极微生物燃料电池(microbial fuel cell,MFC)进行脱氮和产电机理的研究。研究结果发现,阴极碳氮比、pH值对产电和脱氮效率有明显影响。当MFC的阴极运行条件pH值为8.0,碳氮比为20时,运行时间15 h时,脱氮率高达100%,输出电压为150 mV。上述结果表明,微生物燃料电池运行过程中,细菌降解硝酸根的机理为将硝酸根还原为N2或者直接将其作为自身的营养物质而利用。循环伏安(CV)与扫描电镜(SEM)的结果表明,在微生物燃料电池运行中,副球菌YF1通过接触导电作为产电的电子供体。