The mutagenic assessment of an electronic-cigarette and reference cigarette smoke using the Ames assay in strains TA98 and TA100.

Salmonella typhimurium strains TA98 and TA100 were used to assess the mutagenic potential of the aerosol from a commercially available, rechargeable, closed system electronic-cigarette. Results obtained were compared to those for the mainstream smoke from a Kentucky reference (3R4F) cigarette. Two different test matrices were assessed. Aerosol generated from the e-cigarette was trapped on a Cambridge filter pad, eluted in DMSO and compared to cigarette smoke total particulate matter (TPM), which was generated in the same manner for mutagenicity assessment in the Salmonella assay. Fresh e-cigarette and cigarette smoke aerosols were generated on the Vitrocell® VC 10 smoking robot and compared using a modified scaled-down 35mm air agar interface (AAI) methodology. E-cigarette aerosol collected matter (ACM) was found to be non-mutagenic in the 85mm plate incorporation Ames assay in strains TA98 and TA100 conducted in accordance with OECD 471, when tested up to 2400µg/plate. Freshly generated e-cigarette aerosol was also found to be negative in both strains after an AAI aerosol exposure, when tested up to a 1L/min dilution for up to 3h. Positive control responses were observed in both strains, using benzo[a]pyrene, 2-nitrofluorene, sodium azide and 2-aminoanthracene in TA98 and TA100 in the presence and absence of metabolic activation respectively. In contrast, cigarette smoke TPM and aerosol from 3R4F reference cigarettes were found to be mutagenic in both tester strains, under comparable test conditions to that of e-cigarette exposure. Limited information exists on the mutagenic activity of captured e-cigarette particulates and whole aerosol AAI approaches. With the lower toxicant burden of e-cigarette aerosols compared to cigarette smoke, it is clear that a more comprehensive Ames package of data should be generated when assessing e-cigarettes, consisting of the standard OECD-five, TA98, TA100, TA1535, TA1537 (or TA97) and E. coli (or TA102). In addition, TA104 which is more sensitive to the carbonyl based compounds found in e-cigarette aerosols under dry-wicking conditions may also prove a useful addition in a testing battery. Regulatory standard product testing approaches as used in this study will become important when determining whether e-cigarette aerosols are in fact less biologically active than cigarette smoke, as this study suggests. Future studies should be supported by in vitro dosimetry approaches to draw more accurate comparisons between cigarette smoke, e-cigarette aerosol exposure and human use.

The in vitro cytotoxicity and genotoxicity of cigarette smoke particulate matter with reduced toxicant yields.

Tobacco smoke contains more than 5600 constituents, of which approximately 150 are toxicants. This paper describes the activities in the Neutral Red uptake (NRU) assay, the Salmonella mutagenicity test (SAL), the mouse lymphoma mammalian cell mutation assay (MLA) and the in vitro micronucleus test (IVMNT) of Particulate Matter (PM) obtained from experimental cigarettes (ECs), designed to produce reduced levels of toxicants. The designs included tobacco substitute sheet (TSS) containing glycerol, which dilutes toxicants in smoke, or the incorporation of blend-treated (BT) tobacco to reduce the levels of nitrogenous toxicant precursors and some polyphenols. All samples were cytotoxic in the NRU, however TSS reduced PM cytotoxicity in this assay. All PMs were mutagenic in the SAL, MLA and IVMNT. Reductions in bacterial mutagenicity were observed in the SAL, for cigarettes with BT tobacco, compared with their respective controls. The quantitative changes in bacterial mutagenicity could be explained by analytical chemistry data on smoke generated from the ECs used in the study. These observations, and the absence of consistent qualitative differences in the activities of the experimental, control and reference cigarettes, suggest that reduced toxicity cigarettes, as measured by the tests described in this paper, may be developed without introducing any additional cytotoxic or genotoxic hazards, but the impact of this on human health risks remains unknown.

The resolving power of in vitro genotoxicity assays for cigarette smoke particulate matter.

In vitro genotoxicity assays are often used to compare tobacco smoke particulate matter (PM) from different cigarettes. The quantitative aspect of the comparisons requires appropriate statistical methods and replication levels, to support the interpretation in terms of power and significance. This paper recommends a uniform statistical analysis for the Ames test, mouse lymphoma mammalian cell mutation assay (MLA) and the in vitro micronucleus test (IVMNT); involving a hierarchical decision process with respect to slope, fixed effect and single dose comparisons. With these methods, replication levels of 5 (Ames test TA98), 4 (Ames test TA100), 10 (Ames test TA1537), 6 (MLA) and 4 (IVMNT) resolved a 30% difference in PM genotoxicity.

The effect of long term storage on tobacco smoke particulate matter in in vitro genotoxicity and cytotoxicity assays.

Particulate matter (PM) collected from mainstream tobacco smoke is a test article commonly used for in vitro genotoxicity and cytotoxicity testing of combustible tobacco products. However, little published data exists concerning the stability of PM. We completed a 2 year study to quantify the effect of PM storage at -80 °C, on the genotoxicity and cytotoxicity of PM generated from 3R4F and M4A reference cigarettes. The Ames test, Micronucleus assay (MNvit), Mouse Lymphoma assay (MLA) and the Neutral Red Uptake assay (NRU) were used. The majority of M4A and 3R4F PMs were genotoxic and cytotoxic at the timepoints tested. Some minor but statistically significant differences were observed for stored versus freshly prepared PM, but the magnitude of changes were within the variability observed for repeat testing.

The Charles River "hairless" rat mutation maps to chromosome 1: allelic with fuzzy and a likely orthologue of mouse frizzy.

Recent evidence has indicated that the recessive mutation affecting hypotrichosis in the Charles River (CR) "hairless" rat does not involve the hairless gene (hr) on rat chromosome 15. To determine if this mutation might be allelic (or orthologous) with any other previously mapped hypotrichosis-generating mutation in mammals, we have produced a panel of backcross rats segregating for the CR hairless rat mutation as well as numerous other markers from throughout the rat genome. Analysis of this panel has located the CR hairless rat's hypotrichosis-generating mutation on chromosome 1, near Myl2, where only the fuzzy mutation in rat (fz) and the frizzy mutation in mouse (fr) have been previously localized. Intercrossing fz/fz and CR hairless rats produced hybrid offspring with abnormal hair, showing that these two rat mutations are allelic. We suggest that the CR hairless rat mutation and fuzzy be renamed frizzy-Charles River (fr(CR)) and frizzy-Harlan (fr(H)), respectively, to reflect their likely orthology with the mouse fr mutation.

The lecturer-practitioner role in mental health.

The lecturer-practitioner role can be a means of supporting students and making education relevant to practice. The professional credibility--and skills--of the post-holder can be enhanced by this link with practice. It is vital to ensure that the clinical aspects of the post are not neglected.

Imaging of HDR brachytherapy dose distributions using NMR Fricke-gelatin dosimetry.

The application of NMR Fricke-gelatin dosimetry to the imaging of dose distributions around high dose rate (HDR) radioactive Ir-192 seeds in brachytherapy is presented. The duration of the irradiations required to give changes of the Fricke-gelatin NMR characteristics, which were readily detected by the magnetic resonance imager, typically ranged from 4 to 25 min for single source and multiple source (33 source positions distributed among three catheters) geometries, respectively. A fast approach for determining radiation dose from MR image intensity using a calibration curve is described. NMR Fricke-gel dosimetry is shown to be a viable means of verifying the dose distributions from brachytherapy sources.

A novel algorithm for the edge detection and edge enhancement of medical images.

A novel algorithm, histogram shifting (HS) is presented, which performs edge detection or edge enhancement with the choice of two parameters. The histogram of a region surrounding each pixel is found and translated toward the origin, resulting in the new pixel value. Images from a variety of medical imaging modalities were processed with HS to perform detection and enhancement of edges. Comparison with results obtained from conventional edge detection (e.g., Sobel) and with conventional edge-enhancement algorithms is discussed. HS appears to perform the edge-detection operation without introducing "double-edge" effects often obtained with conventional edge-detection algorithms. HS also appears to perform edge enhancement without introducing extensive noise artifacts, which may be noticeable with many edge-enhancement algorithms.

Contrast enhancement of portal images by selective histogram equalization.

Because of the high energy of the treatment beam, contrast of portal verification films is very poor. A simple contrast enhancement technique is described which we have labeled selective histogram equalization (SHE), to improve visualization of double-exposure portal images and this facilitate the beam verification process. The technique performs separate histogram equalization on the treatment- and open-field sections of double-exposure portal images. Delineation of the treatment field edge and separation into two regions is performed automatically for off-line portal radiographs by a strategic combination of Sobel filtration and morphological processes. Analyses of images processed by SHE and other adaptive histogram equalization techniques indicate that SHE produces improved contrast enhancement with minimal addition of noise or artifacts, thus simplifying the beam verification procedure. The simple implementation of an automatic SHE process with on-line portal systems is also discussed.

Teaching nurses therapeutic conversation: a pilot study.

A 2-day video-based workshop developed for teaching psychiatrists was used to instruct learner psychiatric nurses in the conversational model of psychotherapy. A small evaluation was carried out to assess the effects of the teaching on the performance of these nurses in interviewing other learner nurses who role-played people with common emotional difficulties. A randomized groups design was used comparing the interview behaviour of experimental and control groups before and after the workshops, and at 3 months follow-up. Considered as a pilot the study was instructive, although only one significant result emerged. The conversational model of psychotherapy and the teaching workshops devised and used previously with psychiatrists can be used by nurses, including those still in training.