Iturin A-like lipopeptides from Bacillus subtilis trigger apoptosis,paraptosis, and autophagy in Caco-2 cells

This study revealed that iturin A-like lipopeptides produced by Bacillus subtillis induced both paraptosis and apoptosis in heterogeneous human epithelial colorectal adenocarcinoma (Caco-2) cells. Autophagy was simultaneously induced in Caco-2 cells treated with iturin A-like lipopeptides at the early stage and inhibited at the later stage. A western blot analysis showed that the lipopeptides induced apoptosis in Caco-2 cells via a mitochondrial-dependent pathway, as indicated by upregulated expression of the apoptotic genes bax and bad and downregulated expression of the antiapoptotic gene bcl-2. The induction of paraptosis in Caco-2 cells was indicated by the occurrence of many cytoplasmic vacuoles accompanied by endoplasmic reticulum (ER) dilatation and mitochondrial swelling and dysfunction. ER stress also occurred with significant increases in reactive oxygen species and Ca²⁺ levels in cells. Autophagy was detected by a transmission electron microscopy analysis and by upregulated expression of LC3-II and downregulated expression of LC3-I. The inhibition of autophagy at the later stage was shown by upregulated expression of p62. This study revealed the capability of iturin A-like B. subtilis lipopeptides to simultaneously execute antitumor potential via multiple pathways.     

Effect of soluble nickel on cellular energy metabolism in A549 cells

Iron is an essential nutrient to most organisms, and is actively involved in oxygen delivery, electron transport, DNA synthesis, and many other biochemical reactions important for cell survival. We previously reported that nickel (Ni) ion exposure decreases cellular iron level and converts cytosolic aconitase (c-aconitase) to iron-regulatory protein-1 in A549 cells (Chen H, Davidson T, Singleton S, Garrick MD, Costa M. Toxicol Appl Pharmacol 206:275-287, 2005). Here, we further investigated the effect of Ni ion exposure on the activity of mitochondrial iron-sulfur (Fe-S) enzymes and cellular energy metabolism. We found that acute Ni ion treatment up to 1 mM exhibits minimal toxicity in A549 cells. Ni ion treatment decreases the activity of several Fe-S enzymes related to cellular energy metabolism, including mitochondrial aconitase (m-aconitase), succinate dehydrogenase (SDH), and NADH:ubiquinone oxidoreductase (complex I). Low doses of Ni ion for 4 weeks resulted in an increased cellular glycolysis and NADH to NAD+ (NADH/NAD+) ratio, although glycolysis was inhibited at higher levels. Collectively, our results show that Ni ions decrease the activity of cellular iron (Fe)-containing enzymes, inhibit oxidative phosphorylation (OxPhos), and increase cellular glycolytic activity. Since increased glycolysis is one of the fundamental alterations of energy metabolism in cancer cells (the Warburg effect), the inhibition of Fe-S enzymes and subsequent changes in cellular energy metabolism caused by Ni ions may play an important role in Ni carcinogenesis.     

Germ cell specific expression of Vasa in rare minnow, Gobiocypris rarus

Germ cells are set aside early with somatic cells and take roles for reproduction of species from one generation to the next generation. Vasa, a member of DEAD family is well documented as germ cell marker in the animal kingdom. Rare minnow, Gobiocypris rarus, is an emerging model fish in China to study development and toxicology, etc. A suitable germ cell marker will benefit the studies of the factors that may influence germ cell development. Here, we report the cloning and characterization of G. rarus vasa named Grvas whose protein product has the typical characteristics of Vasa proteins. RT-PCR results showed that Grvas is expressed specifically in the gonads of male and female, it is maternally deposited into the eggs for embryos and is continuously expressed in the embryos from the zygote to larvae and adult. Grvas mRNA and/or protein is restricted to the germ cells of ovary and testis. Temporal expression of Grvas mRNA is similar to that of zebrafish vasa during embryogenesis. Grvas signals are coincident with primordial germ cells. These results mean that a germ cell marker, Grvas is isolated from rare minnow and its expression is exclusively in germ cells.     

Genetic homogeneity between populations of cotton bollworm from Xinjiang,China

We studied the population structure of cotton bollworm (CBW), Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), in Xinjiang, the largest cotton-growing region in China, using a fragment of cytochrome c oxidase subunit I (COI) gene. Alignments of all 192 COI sequences revealed 28 haplotypes including 23 in southern Xinjiang, 5 in eastern Xinjiang and 13 in northern Xinjiang. Negative and significant values of neutrality tests for the Tajima's D and Fu's FS parameters, combined with the high values of haplotype diversity (H_d), low values of nucleotide diversity (π) and a high number of low frequency haplotypes indicated a recent demographic expansion of Xinjiang CBW populations. Analysis of molecular variation (AMOVA) indicated low and non-significant genetic structure, regardless of geographical scale or crop, with most of genetic variation occurring within local CBW populations. Pairwise F_ST analyses also indicated low genetic differentiation. This demographic event and high gene flow could be responsible for the low genetic structure currently found. CBW populations in Xinjiang need to be considered as one panmictic unit in its management, especially for the design of refuges to delay the development of resistance by this migratory pest to transgenic Bt (Bacillus thuringiensis) cotton.     

Altered iron homeostasis involvement in arsenite-mediated cell transformation

Chronic exposure to low doses of arsenite causes transformation of human osteogenic sarcoma (HOS) cells. Although oxidative stress is considered important in arsenite-induced cell transformation, the molecular and cellular mechanisms by which arsenite transforms human cells are still unknown. In the present study, we investigated whether altered iron homeostasis, known to affect cellular oxidative stress, can contribute to the arsenite-mediated cell transformation. Using arsenite-induced HOS cell transformation as a model, it was found that total iron levels are significantly higher in transformed HOS cells in comparison to parental control HOS cells. Under normal iron metabolism conditions, iron homeostasis is tightly controlled by inverse regulation of ferritin and transferrin receptor (TfR) through iron regulatory proteins (IRP). Increased iron levels in arsenite transformed cells should theoretically lead to higher ferritin and lower TfR in these cells than in controls. However, the results showed that both ferritin and TfR are decreased, apparently through two different mechanisms. A lower ferritin level in cytoplasm was due to the decreased mRNA in the arsenite-transformed HOS cells, while the decline in TfR was due to a lowered IRP-binding activity. By challenging cells with iron, it was further established that arsenite-transformed HOS cells are less responsive to iron treatment than control HOS cells, which allows accumulation of iron in the transformed cells, as exemplified by significantly lower ferritin induction. On the other hand, caffeic acid phenethyl ester (CAPE), an antioxidant previously shown to suppress As-mediated cell transformation, prevents As-mediated ferritin depletion. In conclusion, our results suggest that altered iron homeostasis contributes to arsenite-induced oxidative stress and, thus, may be involved in arsenite-mediated cell transformation.     

Ultrasound-Assisted Extraction of Water-Soluble Polysaccharides from the Fruit of Acanthopanaxbrachypus: Physicochemical,Structural and Functional Properties

The ultrasound-assisted extraction (UAE) parameters of total water-soluble polysaccharides (TABPs) from Acanthopanaxbrachypus fruit were optimized by Box-Behnken design (BBD) and response surface methodology (RSM). Physicochemical, structural, and functional properties of TABPs were investigated by chemical analysis, inductively coupled plasma optical emission spectrometry (ICP-OES), high performance liquid chromatography (HPLC), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA), water-holding capacity (WHC), oil-holding capacity (OHC), emulsion capacity (EC), emulsion stability (ES), as well as DPPH^. and ABTS^.+scavenging assays. The results showed that the maximal UAE-yield of TABPs was 3.81±0.18 % under the optimal conditions (ultrasonic power 325 W, extraction temperature 47 °C, extraction time 22 min). TABPs was rich in some beneficial element (Mg, K, Fe, Zn and Na) but little in harmful elements (Hg, Cd, As and Pb), and displayed rough surface with flake-like features and large dents, contained 93.89±0.08 % of total carbohydrate with more different monosaccharides including glucose, galactose, rhamnose, arabinose, mannose, xylose, and uronic acid in a molar ratio of 8.83 : 7.90 : 4.74 : 4.55 : 2.80 : 2.39 : 1.00, respectively. TABPs exhibited broad weight distribution (11.2–133.5 kDa), excellent thermal stability (>280 °C), WHC (0.61±0.08 g water/g sample) and OHC (4.53±0.12 g oil/g sample), as well as higher EC (43.75±1.23 %) and ES (38.32±1.50 %). Furthermore, TABPs also displayed remarkable scavenging activities on DPPH^. and ABTS^.+ in vitro. These findings provide a scientific basis for the applications of TABPs in functional additives for food, medicine, and cosmetics.     

Specific expression of Olpiwi1 and Olpiwi2 in medaka (Oryzias latipes) germ cells

Piwi is necessary for germ stem cell survival in Drosophila and homologues have been identified in a diverse range of organisms. Here, we identify and characterize two homologous genes of piwi, Olpiwi1 and Olpiwi2, in the model fish medaka (Oryzias latipes). Olpiwi1 is similar to Ziwi in zebrafish or Miwi in the mouse, and Olpiwi2 is similar to Zili in zebrafish or Mili in the mouse. Moreover, Olpiwi2 mRNA is produced from two different chromosomes. RT-PCR showed expression of Olpiwi1 and Olpiwi2 predominantly in the gonads. In situ hybridization revealed germ cell-specific expression of Olpiwi1 and Olpiwi2 throughout the development of oocytes from oogonia to mature oocytes in the ovary, and from spermatogonia to spermatocytes in the testes of adults. RT-PCR and whole mount in situ hybridization showed that both Olpiwi1 and Olpiwi2 were maternally deposited in the embryo. Olpiwi1 and Olpiwi2 were detected in primordial germ cells during embryonic development. These results suggest that both Olpiwi1 and Olpiwi2 are germ cell specific, and may play important roles in germ cell development and gametogenesis in this model species.