Single walled carbon nanotube reactivity and cytotoxicity following extended aqueous exposure.

Globally carbon nanoparticles are increasingly utilized, yet it is not known if these nanoparticles pose a threat to the environment or human health. This investigation examined 'as-prepared', and acid cleaned carbon nanoparticle physicochemical characteristics (by FTIR, TEM, FESEM, UV-VIS and X-ray microanalysis), and whether these characteristics changed following 2.5-7yr exposure to pH neutral saline or fresh water. To determine if these aqueous aged nanotubes were cytotoxic, these nanotubes were incubated with human epithelial monolayers and analyzed for cell viability (vital staining) and ultrastructural nanoparticle binding/localization (TEM, FESEM). The presence of Ni and Y catalyst, was less damaging to cells than CNT lattice surface oxidation. Extended fresh water storage of oxidized CNTs did not reduce surface reactive groups, nor lessen cell membrane destruction or cell death. However storing oxidized CNTs in saline or NOM significantly reduced CNT-induced cell membrane damage and increased cell survival to control levels.

The exosporium of B. cereus contains a binding site for gC1qR/p33: implication in spore attachment and/or entry.

B. cereus, is a member of a genus of aerobic, gram-positive, spore-forming rod-like bacilli, which includes the deadly, B. anthracis. Preliminary experiments have shown that gC1qR binds to B. cereus spores that have been attached to microtiter plates. The present studies were therefore undertaken, to examine if cell surface gC1qR plays a role in B. cereus spore attachment and/or entry. Monolayers of human colon carcinoma (Caco-2) and lung cells were grown to confluency on 6 mm coverslips in shell vials with gentle swirling in a shaker incubator. Then, 2 microl of a suspension of strain SB460 B. cereus spores (3x10(8)/ml, in sterile water), were added and incubated (1-4 h; 36 degrees C) in the presence or absence of anti-gC1qR mAb-carbon nanoloops. Examination of these cells by EM revealed that: (1) When B. cereus endospores contacted the apical Caco-2 cell surface, or lung cells, gC1qR was simultaneously detectable, indicating upregulation of the molecule. (2) In areas showing spore contact with the cell surface, gC1qR expression was often adjacent to the spores in association with microvilli (Caco-2 cells) or cytoskeletal projections (lung cells). (3) Furthermore, the exosporia of the activated and germinating spores were often decorated with mAb-nanoloops. These observations were further corroborated by experiments in which B.cereus spores were readily taken up by monocytes and neutrophils, and this uptake was partially inhibited by mAb 60.11, which recognizes the C1q binding site on gC1qR. Taken together, the data suggest a role, for gC1qR at least in the initial stages of spore attachment and/or entry.

High resolution FESEM and TEM reveal bacterial spore attachment.

Transmission electron microscopy (TEM) studies in the 1960s and early 1970s using conventional thin section and freeze fracture methodologies revealed ultrastructural bacterial spore appendages. However, the limited technology at that time necessitated the time-consuming process of imaging serial sections and reconstructing each structure. Consequently, the distribution and function of these appendages and their possible role in colonization or pathogenesis remained unknown. By combining high resolution field emission electron microscopy with TEM images of identical bacterial spore preparations, we have been able to obtain images of intact and sectioned Bacillus and Clostridial spores to clearly visualize the appearance, distribution, resistance (to trypsin, chloramphenicol, and heat), and participation of these structures to facilitate attachment of the spores to glass, agar, and human cell substrates. Current user-friendly commercial field emission scanning electron microscopes (FESEMs), permit high resolution imaging, with high brightness guns at lower accelerating voltages for beam sensitive intact biological samples, providing surface images at TEM magnifications for making direct comparisons. For the first time, attachment structures used by pathogenic, environmental, and thermophile bacterial spores could be readily visualized on intact spores to reveal how specific appendages and outer spore coats participated in spore attachment, colonization, and invasion.