Physical and chemical factors influencing the germination of Clostridium difficile spores

AIMS: To investigate the influence of chemical and physical factors on the rate and extent of germination of Clostridium difficile spores. METHODS AND RESULTS: Germination of C. difficile spores following exposure to chemical and physical germinants was measured by loss of either heat or ethanol resistance. Sodium taurocholate and chenodeoxycholate initiated germination together with thioglycollate medium at concentrations of 0.1-100 mmol l(-1) and 10-100 mmol l(-1) respectively. Glycine (0.2% w/v) was a co-factor required for germination with sodium taurocholate. There was no significant difference in the rate of germination of C. difficile spores in aerobic and anaerobic conditions (P > 0.05) however, the initial rate of germination was significantly increased at 37 degrees C compared to 20 degrees C (P < 0.05). The optimum pH range for germination was 6.5-7.5, with a decreased rate and extent of germination occurring at pH 5.5 and 8.5. CONCLUSIONS: This study demonstrates that sodium taurocholate and chenodeoxycholate initiate germination of C. difficile spores and is concentration dependant. Temperature and pH influence the rate and extent of germination. SIGNIFICANCE AND IMPACT OF THE STUDY: This manuscript enhances the knowledge of the factors influencing the germination of C. difficile spores. This may be applied to the development of potential novel strategies for the prevention of C. difficile infection.

Investigation of aspects of reactive gliosis in human astrocytoma cell lines:application for toxicity assessment

The process of astrogliosis, or reactive gliosis, is a typical response of astrocytes to a wide range of physical and chemical injuries. The up-regulation of the astrocyte specific glial fibrillary acidic protein (GFAP) is a hallmark of reactive gliosis and is widely used as a marker to identify the response. In order to develop a reliable, sensitive and high throughput astrocyte toxicity assay that is more relevant to the human response than existing animal cell based models, the U251-MG, U373-MG and CCF-STTG 1 human astrocytoma cell lines were investigated for their ability to exhibit reactive-like changes following exposure to ethanol, chloroquine diphosphate, trimethyltin chloride and acrylamide. Cytotoxicity analysis showed that the astrocytic cells were generally more resistant to the cytotoxic effects of the agents than the SH-SY5Y neuroblastoma cells. Retinoic acid induced differentiation of the SH-SY5Y line was also seen to confer some degree of resistance to toxicant exposure, particularly in the case of ethanol. Using a cell based ELISA for GFAP together with concurrent assays for metabolic activity and cell number, each of the three cell lines responded to toxicant exposure by an increase in GFAP immunoreactivity (GFAP-IR), or by increased metabolic activity. Ethanol, chloroquine diphosphate, trimethyltin chloride and bacterial lipopolysaccharide all induced either GFAP or MTT increases depending upon the cell line, dose and exposure time. Preliminary investigations of additional aspects of astrocytic injury indicated that IL-6, but not TNF-α. or nitric oxide, is released following exposure to each of the compounds, with the exception of acrylamide. It is clear that these human astrocytoma cell lines are capable of responding to toxicant exposure in a manner typical of reactive gliosis and are therefore a valuable cellular model in the assessment of in vitro neurotoxicity.

The influence of environment on foliose lichen growth and its ecological significance

This chapter considers various aspects of the influence of the environment on the growth of foliose lichens and its significance in determining the ecology of individual species. Radial growth (RaG) and growth in mass of foliose lichens is influenced by climate and microclimate and also by substratum factors such as rock and bark texture, substrate chemistry, and nutrient enrichment. Seasonal fluctuations in growth, as measured by radial growth rate (RaGR) per month, often correlate best with average or total rainfall, the number of rain days, or rainfall in a specific season. Temperature has also been identified to be an important climatic factor influencing growth in some studies. Interactions between microclimatic factors and especially light intensity, temperature, and moisture status are important in determining differences in growth in relation to aspect and slope of the substratum. The physical and chemical nature of the substratum has a profound influence on the growth of foliose lichens. Hence, the effects of texture, porosity, rate of drying, and the physical changes of the substratum on growth are likely to influence lichen distributions. Bird droppings may influence growth and survival by smothering the thalli, altering the pH, or adding inhibitory and stimulatory compounds. Nitrogen and phosphate availability may also influence growth. Chemical factors also have an important influence on lichens of maritime rocks, the effect of salinity and calcium ions being of particular importance. Effects of environmental factors on growth influence the competitive ability of a lichen and ultimately its ecology and distribution.

Expert report writing in toxicology:forensic, scientific and legal aspects

Every year throughout the world, individuals' health is damaged by their exposure to toxic chemicals at work. In most cases these problems will resolve, but many will sustain permanent damage. Whilst any justified claim for compensation requires medical and legal evidence a crucial and often controversial component of this process is the establishment of a causal link between the individual's condition and exposure to a specific chemical or substance. Causation, in terms of how a substance or substances led the claimant to his or her current plight, can be difficult to establish and the main purpose of this book, is to provide the aspiring expert report writer with a concise, practical guide that uses case histories to illuminate the process of establishing causation in occupational toxicity proceedings. In summary: A practical, accessible guide to the preparation of balanced, scientifically sound expert reports in the context of occupational toxicology. Focuses on the scientist's role in establishing a causal link between exposure to toxins and an individual's ill health. Includes real-life case histories drawn from the Author's 15 years experience in this area to illustrate the principles involved. Expert Report Writing in Toxicology: Forensic, Scientific and Legal Aspects proves invaluable to scientists across a range of disciplines needing guidance as to what is expected of them in terms of the best use of their expertise and how to present their findings in a manner that is authoritative, balanced and informative.