Peroxisomal membrane channel Pxmp2 in the mammary fat pad is essential for stromal lipid homeostasis and for development of mammary gland epithelium in mice

To understand the functional role of the peroxisomal membrane channel Pxmp2, mice with a targeted disruption of the Pxmp2 gene were generated. These mice were viable, grew and bred normally. However, Pxmp2^−/− female mice were unable to nurse their pups. Lactating mammary gland epithelium displayed secretory lipid droplets and milk proteins, but the size of the ductal system was greatly reduced. Examination of mammary gland development revealed that retarded mammary ductal outgrowth was due to reduced proliferation of epithelial cells during puberty. Transplantation experiments established the Pxmp2^−/− mammary stroma as a tissue responsible for suppression of epithelial growth. Morphological and biochemical examination confirmed the presence of peroxisomes in the mammary fat pad adipocytes, and functional Pxmp2 was detected in the stroma of wild-type mammary glands. Deletion of Pxmp2 led to an elevation in the expression of peroxisomal proteins in the mammary fat pad but not in liver or kidney of transgenic mice. Lipidomics of Pxmp2^−/−mammary fat pad showed a decrease in the content of myristic acid (C₁₄), a principal substrate for protein myristoylation and a potential peroxisomal β-oxidation product. Analysis of complex lipids revealed a reduced concentration of a variety of diacylglycerols and phospholipids containing mostly polyunsaturated fatty acids that may be caused by activation of lipid peroxidation. However, an antioxidant-containing diet did not stimulate mammary epithelial proliferation in Pxmp2^−/− mice.     

Importance of cytochrome c redox state for ceramide-induced apoptosis of human mammary adenocarcinoma cells

Background

Ceramides are intracellular lipid mediator implicated in various cellular responses, including oxidative stress and programmed cell death. Studies demonstrated strong links between ceramide and the mitochondria in the regulation of apoptosis. However, the mechanism of apoptosis induced by ceramides is not fully understood. The present study delineates importance of the redox state of cytochrome c for release of cytochrome c and apoptosis of human mammary adenocarcinoma MCF-7 and MDA-MB-231 cells induced by ceramides.

Methods

The study uses MCF-7 and MDA-MB-231 cells, isolated mitochondria, submitochondrial particles, and oxidized and reduced cytochrome c. Methods used include flow cytometry, immunoblotting, spectroscopy, and respirometry.

Results

We show that ceramides induce mitochondrial oxidative stress and release of cytochrome c from the mitochondria of these cells. Our findings show that ceramides react with oxidized cytochrome c whereas reduced cytochrome c does not react with ceramides. We also show that oxidized cytochrome c reacted with ceramides exerts lower reducibility and function to support mitochondrial respiration. Furthermore, our data show that glutathione protects cytochrome c of reacting with ceramides by increasing the reduced state of cytochrome c.

Conclusions

Ceramides induce oxidative stress and apoptosis in human mammary adenocarcinoma cells by interacting with oxidized cytochrome c leading to the release of cytochrome c from the mitochondria. Our findings suggest a novel mechanism for protective role of glutathione.

General significance

Our study suggests that the redox state of cytochrome c is important in oxidative stress and apoptosis induced by ceramides.     

Evaluation and Spatial Diffusion of Health Risk of Persistent Organic Pollutants (POPs) in Soils Surrounding Chemical Industrial Parks in China

A case study of the cancer risk to humans posed by persistent organic pollutants (POPs) in an industrial area of China, which has a long history of contamination from many sources, is presented. Relatively great concentrations of POPs around the chemical industrial parks have the potential to be chronically carcinogenic to local people. Sixteen individual PAHs listed for priory control by the U.S. Environmental Protection Agency (USEPA), metabolites of DDTs, and isomers of HCHs were measured in soils and a human health risk assessment was conducted by use of USEPA exposure models for children and adults, respectively. Geostatistical methods were used to simulate the spatial diffusion of potential carcinogenic risk, and non-parametric Mann-Whitney U and Kruskal-Wallis tests were employed to analyze the impact of point sources on the surrounding area. The mean value of the sum of Excess Lifetime Cancer Risk (∑ELCR) exceeded the generally acceptable risk level of 1.0E-06 recommended by the USEPA for carcinogenic chemicals. The maximum ∑ELCR was 2.9E-04 for children, which was observed inside the chemical industrial parks. Contamination at the chemical industrial parks caused significant spatial diffusion of ELCR values caused by PAHs, DDT, and HCH.     

Production and characterization of an intracellular bioflocculant by Chryseobacterium daeguense W6 cultured in low nutrition medium

A novel intracellular bioflocculant (named MBF-W6) produced by Chryseobacterium daeguense W6 cultured in low nutrition medium was investigated in this study. The effects of carbon source, nitrogen source, C/N ratio, initial pH, inoculum size, culture temperature and shaking speed on MBF-W6 production were studied. Chemical analysis showed that the purified MBF-W6 was mainly composed of 32.4% protein, 13.1% polysaccharide and 6.8% nucleic acid. Fourier-transform infrared (FTIR) spectroscopy indicated the presence of carboxyl, hydroxyl, and methoxyl groups. The elemental analysis of purified MBF-W6 revealed that the mass proportion of C, H, O, N and S was 40.92:6.53:44.01:8.53:1.01 (w/w) correspondingly. MBF-W6 had good flocculating rate in Kaolin suspension without any cation addition. The highest flocculating rate of 96.9% was achieved under the optimal conditions (bioflocculant dosage 1.2 mg l⁻¹, pH 5.6 and temperature 15 °C).     

产紫青霉EL-02的分离鉴定及其絮凝活性

摘要：【目的】分离鉴定有絮凝活性真菌,同时对其絮凝活性进行初步研究。【方法】采用梯度稀释、平板划线、18S rDNA检测等方法分离鉴定絮凝活性菌株。通过高速离心、超声破碎、乙醇沉淀、定性试验等方法确定絮凝活性物质性质。【结果】从渤海湾海岸土壤样品中分离筛选出一株有较高絮凝活性的真菌,经鉴定为产紫青霉（Penicillium purpurogenum）,命名为产紫青霉EL-02（P. purpurogenum EL-02）。超声破碎试验证实其絮凝活性主要存在于发酵上清液。根据絮凝活性曲线,确定4 d为积累絮     

秦岭南坡不同海拔林分凋落叶分解特征

采用野外放置凋落物分解袋法,对秦岭南坡林区不同海拔华山松(Pinus armandii)、油松(P.tabulaeformis)、锐齿栎(Quercus aliena var.acuteserrata)和华北落叶松(Larix principis-rupprechtii)凋落叶分解过程中N、P、C、粗脂肪、粗纤维和热值的变化规律进行比较研究.结果表明:(1)处于不同海拔的同一树种新鲜凋落叶的N、P、C、粗脂肪含量及其热值差异不显著(P＞0.05).(2)在一年的分解过程中,凋落叶N、P含量表现出逐渐富集的趋势,其中油松凋落叶N、P富集速度最快,分别达到165.60%和189.94%;凋落叶C、粗脂肪含量和热值、C/N、C/P、粗纤维/N在分解中逐渐下降,粗脂肪释放速率达到50.29%～77.82%.(3)分解一年后,不同海拔同一树种凋落叶N、P、C、C/N、C/P含量仍未表现出显著性差异(P＞0.05),但不同海拔凋落叶粗脂肪分解表现出极显著差异(P＜0.01),其中差距最大的锐齿栎凋落叶低海拔较高海拔粗脂肪释放率高19.38%;不同海拔华北落叶松和锐齿栎凋落叶粗纤维释放速率差异极显著(P＜0.01),而不同海拔油松、华山松凋落叶粗纤维释放无显著差异;处于高低海拔的华北落叶松和锐齿栎凋落叶热值分别为17.12和15.68 kJ·g-1、17.74和13.51 kJ·g-1,表现出极显著差异(P＜0.01),油松、华山松凋落叶表现出显著差异(P＜0.05).研究发现,一年中海拔差异所造成的降水、温度等因素的变化对各树种凋落叶中N、P、C的释放无显著影响,但对凋落叶分解过程中粗脂肪、热值、粗纤维/N的变化影响显著.     

Indolacetic and humic acids induce lateral root development through a concerted plasmalemma and tonoplast H<Superscript>+</Superscript> pumps activation

Increasing evidences have indicated that humic substances can induce plant growth and productivity by functioning as an environmental source of auxinic activity. Here we comparatively evaluate the effects of indole-3-acetic acid (IAA) and humic acids (HA) isolated from two different soils (Inseptsol and Ultisol) and two different organic residues (vermicompost and sewage sludge) on root development and on activities of plasmalemma and tonoplast H⁺ pumps from maize roots. The data show that HA isolated from these different sources as well as low IAA concentrations (10⁻¹⁰ and 10⁻¹⁵ M) improve root growth through a markedly proliferation of lateral roots along with a differential activation not only of the plasmalemma but also of vacuolar H⁺-ATPases and H⁺-pyrophosphatase. Further, the vacuolar H⁺-ATPase had a peak of stimulation in a range from 10⁻⁸ to 10⁻¹⁰ M IAA, whereas the H⁺-pyrophosphatase was sensitive to a much broader range of IAA concentrations from 10⁻³ to 10⁻¹⁵ M. It is proposed a complementary view of the acid growth mechanism in which a concerted activation of the plasmalemma and tonoplast H⁺ pumps plays a key role in the root cell expansion process driven by environment-derived molecules endowed with auxinic activity, such as that of humic substances.     

Does secondary chemistry enable lichens to grow on iron-rich substrates?

Lichen substances are shown to increase or to inhibit the adsorption of Fe at cation exchange sites. The influence on the adsorption strongly differs between individual lichen substances and is different for Fe²⁺ and Fe³⁺. These results add a new biological role to the known functions of lichen secondary metabolites. In an experiment with cellulose filters, which were soaked with acetone solutions of lichen substances and were then incubated with micromolar solutions of FeCl₂ or FeCl₃, many lichen substances were found to increase Fe³⁺ adsorption, whereas others had no effect. Most lichen substances had no effect on Fe²⁺ adsorption, but two were found to reduce and one to increase the level of adsorption. Lichens of Fe-poor and -rich sites contain lichen substances with different adsorption behavior towards Fe²⁺ and Fe³⁺. All the studied lichen substances, which only occur in lichens of Fe-poor sites, turned out to be effective Fe³⁺ adsorbents. Lichens of Fe-bearing rock and slag, however, were found to lack lichen substances, or to contain substances that did not adsorb Fe³⁺ and had no effect on Fe²⁺ adsorption, or thirdly, to contain substances that increased Fe³⁺ adsorption, but decreased Fe²⁺ adsorption. These results suggest that lichen substances do play a significant role in Fe adsorption in lichens and determine their tolerance to excess concentrations of Fe. Notwithstanding the strong correlation between the secondary chemistry of lichen species and their preference for Fe-rich or Fe-poor substrates, the postulated mechanism of temporary Fe adsorption by lichen substances has to be subject of future biochemical research.     

Decrease in CD93 (C1qRp) expression in a human monocyte-like cell line (U937) treated with various apoptosis-inducing chemical substances

Human CD93, a receptor for complement component 1, subcomponent q phagocytosis (C1qRp), has been shown to be selectively expressed by cells of a myeloid lineage and was originally reported to be involved in the C1q-mediated enhancement of phagocytosis in innate and adaptive immune responses. The modulation of CD93 expression has been investigated in various cells, particularly in granulocytes and monocytes . We previously reported that a protein kinase C activator (PKC), phorbol myristate acetate (PMA), effectively up-regulated CD93 expression on several cultured cell lines and that its regulation was mainly controlled by a PKC delta-isoenzyme. However, the expression pattern of CD93 in myeloid cells with apoptotic properties remains poorly understood. In this study, we examined the modulation of CD93 expression on a human monocyte-like cell line (U937) treated with various apoptosis-inducing chemical substances : an RNA-synthesis inhibitor, actinomycin D (ActD); a DNA topoisomerase I inhibitor, camptothecin (CPT); a protein-synthesis inhibitor, cycloheximide (CHX); a DNA topoisomerase II inhibitor, etoposide (EPS); and a DNA-synthesis inhibitor, mitomycin C (MMC). Apoptosis was monitored using two-color flow cytometry with Annexin V and 7-amino actinomycin D (7AAD). The above-mentioned substances sufficiently induced the early and late stages of apoptosis, identified as Annexin V positive (+)/7AAD negative (-) cells and Annexin V positive (+)/7AAD positive (+) cells, respectively, in U937 cells after 6 hr of treatment. The modulation of CD93 expression on U937 cells during the early stage of apoptosis, gated as Annexin V positive (+)/7AAD negative (-) cells, was then investigated using a CD93 mAb (mNI-11), originally established in our laboratories, and flow cytometry using a fluorescence-activated cell sorter (FACS). The mean fluorescence intensity (MFI) of the cells that stained positive for CD93 mAb (mNI-11) among the treated U937 cells showed a dramatic decrease in expression. In addition, the expressions of HLA-class I (HLA-A, B, C), HLA-class II (HLA-DR), CD18 (lymphocyte function-associated antigen-1 beta; LFA-1beta) and CD54 (intercellular adhesion molecule-1; ICAM-1) were also markedly decreased on the treated U937 cells identified as Annexin V positive (+)/7AAD negative (-) cells (early stage of apoptosis). Interestingly, the expression patterns of CD93 on the U937 cells treated with the above-mentioned chemical substances closely resembled those of HLA-class I (HLA-A, B, C). An immunoblotting analysis showed that the expression of a surface antigen (molecular size, about 97 kDa) targeted by the CD93 mAb (mNI-11) on the U937 cells treated with various apoptosis-inducing chemical substances had clearly decreased. On the other hand, an enzyme-linked immunoassay (EIA) showed that although PMA-treated U937 cells had strongly secreted soluble CD93 (sCD93) into the culture supernatant, the secretion of sCD93 in the culture supernatant of the U937 cells treated with the above-mentioned chemical substances was not enhanced, compared with that of untreated U937 cells. Importantly, however , the U937 cells with apoptotic properties induced by various apoptosis-inducing chemical substances also rapidly (in 30 min) and strongly secreted sCD93 into the culture supernatant in the presence of PMA. Taken together, these findings indicate that the expression of the CD93 molecule identified by CD93 mAb (mNI-11) is dramatically decreased on U937 cells with apoptotic properties, and that the decrease in CD93 expression on U937 cells treated with apoptosis-inducing chemical substances may be a good model for analyzing the regulation of CD93 expression on apoptotic myeloid cells.     

水体硝化体系中砷的解毒机制探讨

硝化是目前废水生物脱氮中应用最为广泛的工艺之一,其功能菌为化能自养型细菌,生长缓慢,对重金属十分敏感。砷是一种剧毒的类金属元素,主要以无机形式的亚砷酸盐[AsO₂^-,As（Ⅲ）]和砷酸盐[AsO₄^3-,As（Ⅴ）]存在,尤以As（Ⅲ）毒性最强。但研究发现,在硝化体系中,高浓度As（Ⅲ）(约400mg/L)未对硝化功能微生物产生明显毒性。深入比较发现,As（Ⅲ）的生物氧化与硝化过程具有一定的关联性。化能自养型As（Ⅲ）氧化菌不仅可在有氧条件下将As（Ⅲ）氧化,还可在缺氧条件下以NO₂^–或NO₃^–为电子受体氧化As（Ⅲ）。而硝化细菌也是典型的化能自养菌,且硝化体系内存在氧气及硝化产物NO₂^–、NO₃^–等电子受体,理论上均可接受电子实现As（Ⅲ）的氧化。本文结合硝化反应特性,综述了As（Ⅲ）在硝化体系下的解毒机制,主要为胞外聚合物的...