Histochemical Study of the Progenetic Trematode Alloglossidium renale

A histochemical study of the progenetic trematode Alloglossidium renale has demonstrated the absence of any secreted material between the adult worm and the host (freshwater shrimp) antennal gland tubules. Host tissue is affected only by the compression, abrasion, and ingestion by the parasite, and host tubule cells near the worm have the same staining patterns as those distant from the parasite. The trematode sometimes dies within the host, leaving a necrotic mass whose histochemical staining differs significantly from the living organism. In the necrotic mass, the only recognizable features were the ova and the vitellarium, which atrophied and resulted in tyrosine-positive staining within the mass. A melanin reaction was not observed in the host using a specialized ferro-ferricyanide stain. The only apparent host response to infection was a layer of damaged squamous host cells adhering to the necrotic worm. The results confirm benign host-parasite effects and a highly evolved relationship between the host and parasite, perhaps bordering on commensalism

Meiofauna and Trace Metals From Sediment Collections in Florida After the Deepwater Horizon Oil Spill

Sediment from the Florida Gulf continental shelf was collected from 18 sites during October and November 2010 for meiofauna and trace-metals analysis. Collections were obtained using a Shipek® grab on the National Oceanic and Atmospheric Administration ship Pisces and spanned from the head of the DeSoto Canyon to off the southern end of the Florida peninsula approximately following the 100–200-m contour. Mean abundance of the dominant meiofaunal groups (nematodes, copepods, and polychaetes) was unchanged when compared with 2007–2009 data. Nematodes and copepods correlated positively with each other, and negatively with latitude and longitude, suggesting that there were higher densities in southern Florida. These results contrast with those from 2007–2009 in that previously nematodes had no correlation with latitude or longitude in Florida. Nickel (Ni) and vanadium (V) concentrations were higher in the western Florida locations and correlated positively with increasing depth. No relationship was found between Ni, V, and meiofauna densities

Expression of lung vascular and airway ICAM-1 after exposure to bacterial lipopolysaccharide

Airway instillation of bacterial lipopolysaccharide (LPS) into rat lungs induces neutrophil accumulation, which is known to be intercellular adhesion molecule-1 (ICAM-1)-dependent. In the present study, ICAM-1 messenger RNA (mRNA) of whole lung was found to increase by 20-fold in this inflammatory model. This increase was reduced by 81 % after treatment of animals with anti-tumor necrosis factor- � (TNF-�) antibody and by 37 % after treatment with anti-interleukin-1 (IL-1) antibody. The same interventions reduced whole-lung ICAM-1 protein by 85 % and 25%, respectively. The studies were extended to assess the locale in lung of ICAM-1 upregulation. Lung vascular ICAM-1 content, which was assessed by vascular fixation of [ 125 I]anti-ICAM-1, rose 4-fold after airway instillation of LPS. This rise was also TNF-�dependent. Under the same experimental conditions, fixation of [ 125 I]anti-ICAM-1 to airway surfaces increased 11-fold in a TNF-�-dependent manner. In situ hybridization and immunohistochemical analyses of lung tissue revealed ICAM-1 upregulation in the bronchiolar epithelium and in peribronchiolar smooth muscle. Soluble ICAM-1 could also be detected in bronchoalveolar lavage fluids (BALFs) of animals after intratracheal instillation of LPS. Retrieved alveolar macrophages showed a small, significant, and transient increase in surface expression of ICAM-1. These data indicate, at the very least, a dual compartmentalize

Suppression of cancer progression by MGAT1 shRNA knockdown.

Oncogenic signaling promotes tumor invasion and metastasis, in part, by increasing the expression of tri- and tetra- branched N-glycans. The branched N-glycans bind to galectins forming a multivalent lattice that enhances cell surface residency of growth factor receptors, and focal adhesion turnover. N-acetylglucosaminyltransferase I (MGAT1), the first branching enzyme in the pathway, is required for the addition of all subsequent branches. Here we have introduced MGAT1 shRNA into human HeLa cervical and PC-3-Yellow prostate tumor cells lines, generating cell lines with reduced transcript, enzyme activity and branched N-glycans at the cell surface. MGAT1 knockdown inhibited HeLa cell migration and invasion, but did not alter cell proliferation rates. Swainsonine, an inhibitor of α-mannosidase II immediately downstream of MGAT1, also inhibited cell invasion and was not additive with MGAT1 shRNA, consistent with a common mechanism of action. Focal adhesion and microfilament organization in MGAT1 knockdown cells also indicate a less motile phenotype. In vivo, MGAT1 knockdown in the PC-3-Yellow orthotopic prostate cancer xenograft model significantly decreased primary tumor growth and the incidence of lung metastases. Our results demonstrate that blocking MGAT1 is a potential target for anti-cancer therapy

shRNA MGAT1 suppresses N-glycan branching.

<p><b>A</b>) HeLa cells were infected with lentiviral vectors targeting MGAT1(shRNA1 or shRNA2) or the control shRNA sequences. Stable cell populations were selected by the addition of puromycin (1 µg/mL). <b>A</b>)MGAT1 mRNA were measured by qRT-PCR, <b>B</b>) MGAT1 enzyme activity, <b>C</b>) L-PHA reactive surface N-glycans by Array scan microscope, <b>D</b>) Proliferation over 4 days Data represent the mean ± SD relative expression of mRNA relative to control sequence (n = 3 independent experiments performed in triplicate).</p