Purification of long helical capsid of newcastle disease virus from Escherichia coli using anion exchange chromatography

NP_Δc375 is a truncated version of the nucleocapsid protein of Newcastle disease virus (NDV) which self‐assembles into a long helical structure. A packed bed anion exchange chromatography (PB‐AEC), SepFastTM Supor Q pre‐packed column, was used to purify NP_Δc375 from clarified feedstock. This PB‐AEC column adsorbed 76.2% of NP_Δc375 from the clarified feedstock. About 67.5% of the adsorbed NP_Δc375 was successfully eluted from the column by applying 50 mM Tris‐HCl elution buffer supplemented with 0.5 M NaCl at pH 7. Thus, a recovery yield of 51.4% with a purity of 76.7% which corresponds to a purification factor of 6.5 was achieved in this PB‐AEC operation. Electron microscopic analysis revealed that the helical structure of the NP_Δc375 purified by SepFast^TM Supor Q pre‐packed column was as long as 490 nm and 22–24 nm in diameter. The antigenicity of the purified NP_Δc375 was confirmed by enzyme‐linked immunosorbent assay. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29: 564–567, 2013     

Long-term effects of induced mineral accretion on growth, survival and corallite properties of Porites cylindrica Dana

The mineral accretion technology involves the accumulation of dissolved mineral ions within the vicinity of underwater electrodes and their deposition via electrochemical processes onto the positive electrode forming a natural substrate. Branches of Porites cylindrica were reared under the presence of mineral accretion (treated nubbins), without mineral accretion (untreated), and under undisturbed (control) conditions for 6 months (mineral accretion phase). The electricity for the treated nubbins was cut off after the sixth month, and all remaining nubbins were allowed to grow for another 6 months (post mineral accretion phase). The longitudinal growth rate of the treated nubbins was relatively high during the mineral accretion phase then dropped during the post mineral accretion phase. Statistical analysis revealed longitudinal growth to be significant over time, but the significant differences between treatments lay only between the first until the second bi-monthly period (from January to May 2000) and the rest of the observation period (June to January 2001). This indicates that growth enhancement occurred only during the early stages of the mineral accretion phase. There were also significant differences in girth growth between phases and between treatments as well as a significant phase by treatment interaction again indicating that significant differences between treatments occurred only during a certain phase. In terms of overall survival, the treated nubbins fared better than the untreated nubbins. The corals in all treatments showed the same trend in corallite size and density after 1 year. Corallite sizes increased from the tip towards the base of the transplant. The reverse trend occurred with corallite density which decreased from the tip towards the base. Phenotypic alteration of the corallites (decrease in size and increase in density from the middle region of the nubbin towards the base) had occurred in the treated nubbins while they were exposed to mineral accretion. This effect disappeared, however, after 6 months. It appears, therefore, that enhancement of growth took place only during active mineral accretion, though there were positive, longer-term effects on survival.     

Arachidonic Acid Activation of Monocyte and Neutrophil Reactive Oxygen in Lung Cancer Patients Undergoing Pulmonary Resection

Reactive oxygen (roi) generation was investigated in phagocyted of 39 patients undergoing pulmonary resection for lung cancer and 39 paired healthy controls. Generation of roi in monocytes and neutrophils was monitored using 2′, 7′-dichlorofluorescein diacetate. Activation associated with hydrophobic interactions was probed by analysis of phagocyte roi activation by arachidonic acid and γ-linolenic acid. Patient roi was measured pre-operatively and 2 and 7 days post-operatively. Elevated (P<0.01) roi productin was detected in neutrophils of lung cancer patients. surgery was associated with an increase (P<0.05–P<0.01) in phagocyte roi at 2 and 7 days post-op. Phagocyte roi was stimulated by arachidonic acid and γ-linolenic acid (1–40 μ ) both pre- and post-operatively. Differences in arachidonic acid and γ-linolenic acid stimulation between patient and control and pre- and post-op patient phagocyted suggest arachidonic acid involvement in phagocyte activation during reactive responses to lung carcinoma and surgery.     

Molecular characterization of the Caenorhabditis elegans Rho GDP-dissociation inhibitor.

GDP-dissociation inhibitors (GDIs) form one of the classes of regulatory proteins that modulate the cycling of the Ras superfamily of GTPases between active GTP-bound and inactive GDP-bound states. We report here the characterization of the Caenorhabditis elegans RhoGDI (CeRhoGDI) as part of our investigations into Rho-GTPase signalling pathways that are involved in nematode development. CeRhoGDI is a 23-kDa protein that is localized predominantly in the cytosol. CeRhoGDI interacts only with the lipid-modified forms of C. elegans Rho-GTPases, CeRhoA, CeRac1 and Cdc42Ce, in vitro and is able to solubilize the membrane-bound forms of these GTPases. CeRhoGDI recognizes the GTPases in both GTP- and GDP-bound forms; hence it inhibits both the guanine-nucleotide dissociation and GTP-hydrolysis activities. The inhibitory activity towards the GTP-bound GTPases is weak compared with that towards GDP-bound GTPases. CeRhoGDI is expressed throughout development and is highly expressed in marginal and vulval epithelial cells, in sperm cells and spicules. Taken together, our results suggest that CeRhoGDI may be involved in specific morphogenetic events mediated by the C. elegans Rho-GTPases.     

Some ultrastructural observations on the microfilaria of Breinlia sergenti--nervous system

The amphids, cephalic papillae, phasmids, excretory complex, anal vesicle, rectal cells, G cell and the nerve ring are described in the microfilaria of Breinlia sergenti. The significance of these various structures in playing a nervous role in this organism is discussed, and a working hypothesis presented to explain the nervous co-ordination in this parasitic stage.     

The effect of interleukin-1, interleukin-6 and its interrelationship on the synthesis of serum amyloid A and C-reactive protein in primary cultures of adult human hepatocytes

During the acute phase response, synthesis of C-reactive protein and serum amyloid A is increased. To investigate whether the enhanced synthesis of these proteins are due to stimulatory effect of inflammatory mediators such as interleukin-1 (IL-1) and interleukin-6 (IL-6) produced by macrophages and monocytes, primary cultures of adult human hepatocytes were exposed to recombinant (r)IL-1, rIL-6 or rIL-1 and monospecific anti rIL-6 antibodies in the presence of 1 microM dexamethasone. The findings indicate that rIL-1 and rIL-6 both stimulate the liver synthesis of C-reactive protein and serum amyloid A, however monospecific anti rIL-6 antibodies reduce the stimulatory effect of rIL-1 on the synthesis of these proteins. These findings suggest that IL-6 plays a key role in the stimulation of synthesis of serum amyloid A and C-reactive protein by the human liver cells.