Fungi isolated from packaging materials: Their role in the production of 2, 4, 6-trichloroanisole

Fungi isolated from fibreboard cartons, paper sacks and jute sacks were assessed for their ability to methylate 2, 4, 6-trichlorophenol in a defined, reduced-water-activity growth medium. When extracts of individual cultures were analysed by gas chromatography-multiple ion monitoring mass spectrometry, 17 of the 31 species, initially selected by sensory techniques, were found to produce 2, 4, 6-trichloroanisole. Quantitative analyses showed that the most efficient methylators in the defined liquid medium were Paecilomyces variotii, Fusarium oxysporum, Penicillium crustosum, Pen citrinum, Pen brevicompactum and one strain of Aspergillus flavus. The role of these fungi in the production of 2, 4, 6-trichloroanisole in packaging materials contaminated with 2, 4, 6-trichlorophenol is discussed.     

Raman spectroscopic discrimination of cell response to chemical and physical inactivation

Raman spectroscopy was applied to study Escherichia coli and Staphylococcus epidermidis cells that were inactivated by different chemicals and stress conditions including starvation and high temperature. E. coli cells exposed to starvation conditions over several days lost viability at the same rate that spectral bands assigned to DNA and RNA bases decreased in intensity. Band intensities correlate with standard plate counts with R(2) = 0.99 and R(2) = 0.97, respectively. Principal components analysis and discriminant analysis multivariate statistical techniques were used to evaluate the spectral data collected. Significant changes were observed in the spectra of treated cells in comparison with their respective controls (samples without treatment). As a result, there was a significant differentiation between viable and non-viable cells (treated and non-treated cells) in the first and second principal component plots for all the treatments. Discriminant analysis was used along with PCA to estimate a classification rate based on viability status of the cells. Non-viable cells were differentiated from viable cells with classification rates that ranged between 60 and 90% for specific treatments (i.e., EDTA-treated cells versus control cells). The classification rate obtained considering all the treatments (non-viable cells) and controls (viable cells) at the same time for each of the species studied was 86%. The classification rate based on species differentiation when all the spectra (viable and non-viable) were used was 87%. These results suggest that Raman spectroscopy is a powerful tool that can be used to evaluate viability and to study metabolic changes in microorganisms. It is a robust method for bacterial identification even when high spectral variations are introduced.     

Non-contrast MRI methods as a tool for the pre-operative assessment and surveillance of the arterio-venous fistula for haemodialysis

Objective

To compare non-contrast enhanced MRI with ultrasound (US) for measurement of arm blood vessel geometries and flow velocities in volunteers and patients with end-stage renal disease.

Materials and methods

Subjects were scanned using US (reference standard), and MRI 2D time-of-flight (ToF), 2D phase contrast (PC), and 3D multi-echo data image combination (MEDIC). Patients were also scanned after arteriovenous fistula (AVF) surgery.

Results

For mean vessel diameters (radial and brachial arteries; cephalic vein) MEDIC measurements were similar to US (p?>?0.05). However, ToF underestimated the mean diameter of the cephalic vein relative to US (p?<?0.05). For arterial velocity measurements, the mean values derived by PC-MR and US were similar (p?>?0.05). Post-operatively, the intra-luminal signal intensity was hypo-intense at the anastomosis site using ToF and MEDIC. At the same site the outer boundary of the vessel was consistently lost on ToF, but remained clearly delineated on the MEDIC images.

Discussion

With the exception of ToF, the MRI data demonstrated excellent agreement with US for measurements of vessel geometry and flow velocity. Further, the ability to clearly delineate the post-surgery vessel edges with MEDIC MRI suggests that the technique may be useful for surveillance after AVF creation or for patient-specific modelling studies.

     

A thixotropic nanocomposite gel for three-dimensional cell culture

Thixotropic materials, which become less viscous under stress and return to their original state when stress is removed, have been used to deliver gel-cell constructs and therapeutic agents. Here we show that a polymer-silica nanocomposite thixotropic gel can be used as a three-dimensional cell culture material. The gel liquefies when vortexed--allowing cells and biological components to be added--and resolidifies to trap the components when the shear force from spinning is removed. Good permeability of nutrients and gases through the gel allows various cell types to proliferate and be viable for up to three weeks. Human mesenchymal stem cells cultured in stiffer gels developed bone-like behaviour, showing that the rheological properties of the gel can control cell differentiation. No enzymatic, chemical, or photo-crosslinking, changes in ionic strength or temperature are required to form or liquefy the gel, offering a way to sub-culture cells without using trypsin-a protease commonly used in traditional cell culture techniques.     

Surprising shrinkage of expanding gels under an external load

Hydrogels are fascinating and useful in that they can show large volume changes in response to various stimuli, such as temperature or chemical environment. Here we report the peculiar observation that chemically crosslinked hydrogels that normally expand owing to a change in electrolyte pH can be made to shrink in certain circumstances. Specifically, these hydrogels contract when tested at a constant compressive force and subjected to a pH change that causes expansion in the absence of the applied load. When tested under tension, the gels always expand. Although the effects of external stress on the swelling of gels is known, the concomitant change in gel mechanical properties during pH switching was found to be a more dominant effect in our studies. However, existing mechanical models used to predict dimensional changes in actuator materials could not explain both the tensile and compression results. In addition, we show that the friction between metal plates of the apparatus and the gel is a key factor in explaining the contractile actuation under compressive loads. The observations reported in this paper are important for the successful design and use of hydrogel actuators in devices such as valves for microfluidics.     

Studying bacterial metabolic states using Raman spectroscopy

Natural metabolic variability expected during characteristic growth phases in batch cultures of Escherichia coli and Staphylococcus epidermidis were studied by Raman spectroscopy. Spectral changes induced by metabolic changes found in the growth phases (i.e., lag, exponential, stationary, and decay) were identified. Maximum intensity of bands assigned to DNA and RNA bases are seen at the beginning of the exponential phase, when cells are metabolically active, and minimum intensities are seen when cells are decaying. High agreement in spectral variation due to growth phases was seen for all the trials that were performed, four growth cycles for E. coli and two for S. epidermidis. Batch cultures were monitored by standard plate counts to identify all growth phases, including decay. Spectral data were analyzed by principal component analysis (PCA) and discriminant analysis to identify similarities and differences and to estimate a classification performance based on growth phases. For the species evaluated, spectra during decay are grouped closer to each other and separated from lag, exponential, and stationary cells. These results suggest that Raman spectroscopy can be used to study metabolic states in bacteria and in particular cell viability.