Effect of long-term exposure to simulated acid rain on the development and reproduction of the predatory mite, <Emphasis Type="Italic">Euseius nicholsi</Emphasis> (Ehara <Emphasis Type="Italic">et</Emphasis> Lee) (Acari: Phytoseiidae)

The effects of simulated acid rain on the development and reproduction of E. nicholsi, an important natural enemy of many species of pest mites in China, were studied in the laboratory. The life history parameters of E. nicholsi were investigated after long-term (about 8 generations) exposure to various levels of simulated acid rain (pH 2.0, 3.0, 4.0, and 5.6). The results showed that the acid rain had negative effects on some basic parameters of the life history of the E. nicholsi population indirectly. The whole development durations with pH 3.0–5.6 treatments were longer and the duration with pH 2.0 treatment was shorter than with deionized water treatments (control). Adult longevity and oviposition duration were notably shortened, and female fecundity was decreased by an increase in acidity. The net reproductive rate (R ₀) of the populations treated with acid rain was less than that of the control populations and gradually decreased with an increase in acidity; however, the values of the intrinsic rate of increase (r) were not affected significantly by acid rain, and inhibitory effects of acid rain on the population growth of E. nicholsi were not recognized.     

Cyclase‐associated protein 1 is a key negative regulator of milk synthesis and proliferation of bovine mammary epithelial cells

Adenylyl cyclase‐associated protein (CAP) is a highly conserved protein. Previous reports have suggested that CAP1 may be a negative regulator of cellular proliferation, migration, and adhesion and the development of cell carcinomas. The molecular mechanism of CAP1 regulation of downstream pathways, as well as how CAP1 is regulated by environmental stimuli and upstream signalling, is not well understood. In this present study, we assessed the role of CAP1 in milk synthesis and proliferation of bovine mammary epithelial cells. Using gene overexpression and silencing methods, CAP1 was found to negatively regulate milk synthesis and proliferation of cells via the PI3K‐mTOR/SREBP‐1c/Cyclin D1 signalling pathway. Hormones, such as prolactin and oestrogen, and amino acids, such as methionine and leucine, stimulate MMP9 expression and trigger CAP1 degradation, and thus, abrogate its inhibition of synthesis of milk protein, fat, and lactose by and proliferation of bovine mammary epithelial cells. The results of our study help deepen our understanding of the regulatory mechanisms underlying milk synthesis and aid in characterizing the molecular mechanisms of CAP1. Previous reports have suggested that CAP1 is a negative regulator of cellular proliferation and anabolism, but the molecular mechanisms are largely unknown. In this present study, we identified CAP1 as a negative regulator of milk synthesis and proliferation of bovine mammary epithelial cells. Our results will deepen our understanding of the regulatory mechanisms underlying milk synthesis and aid in exploring the molecular mechanisms of CAP1.     

Phytane and neoclerodane diterpenes from the aerial parts of Inula nervosa Wall.

Three new aliphatic diterpenes (1–3), together with three known neoclerodane-type diterpenes (4–6) were isolated from the aerial parts of Inula nervosa Wall. The structures of 1-3 were elucidated on the basis of 1D and 2D spectroscopic analysis. Additionally, phytane-type and neoclerodane-type diterpenes have not been reported in any species of the genus Inula yet. The phytane-type and neoclerodane-type diterpenes obtained from I. nervosa Wall. suggest this plant maybe have remote genetic relations with other Inula species.     

Reactivation of Horseradish peroxidase in organic solvent using extraction

A novel extraction method was developed aiming at increasing the stability of enzymes in organic solvent media. Horseradish peroxidase (HRP), inactivated in a tetrahydrofuran (THF)/water (1:1, v/v), regained and maintained its activity when HRP was extracted by adding a THF/benzene mixture to the original solution. However, the HRP activity was drastically lowered in the enzyme-free blank solution that had been formed by employing the same extraction procedure. As a result, the reactivation after the extraction is believed to depend on enzyme history, and might be arisen from an irreversible structural change of the enzyme.     

Analysis and characterization of the functional TGFβ receptors required for BMP6-induced osteogenic differentiation of mesenchymal progenitor cells

Although BMP6 is highly capable of inducing osteogenic differentiation of mesenchymal progenitor cells (MPCs), the molecular mechanism involved remains to be fully elucidated. Using dominant negative (dn) mutant form of type I and type II TGFβ receptors, we demonstrated that three dn-type I receptors (dnALK2, dnALK3, dnALK6), and three dn-type II receptors (dnBMPRII, dnActRII, dnActRIIB), effectively diminished BMP6-induced osteogenic differentiation of MPCs. These findings suggested that ALK2, ALK3, ALK6, BMPRII, ActRII and ActRIIB are essential for BMP6-induced osteogenic differentiation of MPCs. However, MPCs in this study do not express ActRIIB. Moreover, RNA interference of ALK2, ALK3, ALK6, BMPRII and ActRII inhibited BMP6-induced osteogenic differentiation in MPCs. Our results strongly suggested that BMP6-induced osteogenic differentiation of MPCs is mediated by its functional TGFβ receptors including ALK2, ALK3, ALK6, BMPRII, and ActRII. [BMB Reports 2013; 46(2): 107-112]     

Precursor engineering and cell physiological regulation for high level rapamycin production by Streptomyces hygroscopicus

A process for high level production of rapamycin by Streptomyces hygroscopicus using statistical designs and feeding strategy was developed. The amino acids (i.e. Lys, Tyr, and Gln) for precursor supply were screened out in the initial phase of fermentation. The optimum levels determined with Box-Behnken design were Lys 20, Tyr 4, and Gln 3 g/l. In the rapamycin biosynthesis phase, the important component, ammonium sulphate, was also identified. A novel two-stage feeding strategy was developed successfully to increase the flux of rapamycin biosynthesis, in which the optimized amino acid components were fed in the initial phase of fermentation, and then switched to feed 2 g/l ammonium sulphate at 72 h. The maximal rapamycin production reached 860.6 mg/l in a 7 l fermentor, which was 182 % higher than that of the control. This was the first report to integrate precursor engineering and cell physiological regulation methods to optimize rapamycin production.     

Ecology of Hantaan virus at Twin Bridges Training Area,Gyeonggi Province,Republic of Korea, 2005–2007

The Twin Bridges Training Area (TBTA) in the Republic of Korea consists of dirt roads, barren training areas, and forested hillsides adjacent to linear and broad expanses of tall grasses, herbaceous, and scrub vegetation. Of the six species of small mammals, the striped field mouse, Apodemus agrarius, was the most frequently captured (96.1%). Apodemus agrarius capture rates varied from 17.7 to 33.2% during three trapping periods. Gravid females were observed during November‐December 2006 (8.4%) and March 2007 (5.1%). In 2005, the overall seroprevalence of Hantaan virus (HTNV) was high (34.4%) and lower during surveys in 2006 (14.2%) and 2007 (13.8%). Seroprevalence was directly correlated with weight increase of A. agrarius.     

Intratumour Diversity of Chromosome Copy Numbers in Neuroblastoma Mediated by On-Going Chromosome Loss from a Polyploid State

Neuroblastomas (NBs) are tumours of the sympathetic nervous system accounting for 8–10% of paediatric cancers. NBs exhibit extensive intertumour genetic heterogeneity, but their extent of intratumour genetic diversity has remained unexplored. We aimed to assess intratumour genetic variation in NBs with a focus on whole chromosome changes and their underlying mechanism. Allelic ratios obtained by SNP-array data from 30 aneuploid primary NBs and NB cell lines were used to quantify the size of clones harbouring specific genomic imbalances. In 13 cases, this was supplemented by fluorescence in situ hybridisation to assess copy number diversity in detail. Computer simulations of different mitotic segregation errors, single cell cloning, analysis of mitotic figures, and time lapse imaging of dividing NB cells were used to infer the most likely mechanism behind intratumour variation in chromosome number. Combined SNP array and FISH analyses showed that all cases exhibited higher inter-cellular copy number variation than non-neoplastic control tissue, with up to 75% of tumour cells showing non-modal chromosome copy numbers. Comparisons of copy number profiles, resulting from simulations of different segregation errors to genomic profiles of 120 NBs indicated that loss of chromosomes from a tetraploid state was more likely than other mechanisms to explain numerical aberrations in NB. This was supported by a high frequency of lagging chromosomes at anaphase and polyploidisation events in growing NB cells. The dynamic nature of numerical aberrations was corroborated further by detecting substantial copy number diversity in cell populations grown from single NB cells. We conclude that aneuploid NBs typically show extensive intratumour chromosome copy number diversity, and that this phenomenon is most likely explained by continuous loss of chromosomes from a polyploid state.