Spatial analysis of biomineralization associated gene expression from the mantle organ of the pearl oyster Pinctada maxima

Background: Biomineralization is a process encompassing all mineral containing tissues produced within an organism. One of the most dynamic examples of this process is the formation of the mollusk shell, comprising a variety of crystal phases and microstructures. The organic component incorporated within the shell is said to dictate this architecture. However general understanding of how this process is achieved remains ambiguous. The mantle is a conserved organ involved in shell formation throughout molluscs. Specifically the mantle is thought to be responsible for secreting the protein component of the shell. This study employs molecular approaches to determine the spatial expression of genes within the mantle tissue to further the elucidation of the shell biomineralization. Results: A microarray platform was custom generated (PmaxArray 1.0) from the pearl oyster Pinctada maxima. PmaxArray 1.0 consists of 4992 expressed sequence tags (ESTs) originating from mantle tissue. This microarray was used to analyze the spatial expression of ESTs throughout the mantle organ. The mantle was dissected into five discrete regions and analyzed for differential gene expression with PmaxArray 1.0. Over 2000 ESTs were determined to be differentially expressed among the tissue sections, identifying five major expression regions. In situ hybridization validated and further localized the expression for a subset of these ESTs. Comparative sequence similarity analysis of these ESTs revealed a number of the transcripts were novel while others showed significant sequence similarities to previously characterized shell related genes.

Socio-economic differences in takeaway food consumption among adults

To examine socioeconomic differences in the frequency and types of takeaway foods consumed. Cross-sectional postal survey. Participants were asked about their usual consumption of overall takeaway food (< four times a month, or ≥ four times a month) and 22 specific takeaway food items (< once a month, or ≥ once a month): these latter foods were grouped into “healthy” and “less healthy” choices. Socioeconomic position was measured using education and equivalised household income and differences in takeaway food consumption were assessed by calculating prevalence ratios using log binomial regression. Adults aged 25–64 years from Brisbane, Australia were randomly selected from the electoral roll (N = 903, 63.7% response rate). Compared with their more educated counterparts, the least educated were more regular consumers of overall takeaway food, fruit/vegetable juice, and less regular consumers of sushi. For the “less healthy” items, the least educated more regularly consumed potato chips, savoury pies, fried chicken, and non-diet soft drinks; however, the least educated were less likely to consume curry. Household income was not associated with overall takeaway consumption. The lowest income group were more regular consumers of fruit/vegetable juice compared with the highest income group. Among the “less healthy” items, the lowest income group were more regular consumers of fried fish, ice-cream, and milk shakes, while curry was consumed less regularly. The frequency and types of takeaway foods consumed by socioeconomically disadvantaged groups may contribute to inequalities in overweight/obesity and chronic disease.

Estimation on the mortality and disease burden attributed to selected risk factors in Shandong province [山东省主要危险因素的归因死亡和疾病负担分析]

Objective: To determine the major health related risk factors and provide evidence for policy-making,using health burden analysis on selected factors among general population from Shandong province. Methods: Based on data derived from the Third Death of Cause Sampling Survey in Shandong. Years of life lcrat(YLLs),yearS Iived with disability(YLDs)and disability-adjusted life years(DALYs) were calculated according to the GBD ethodology.Deaths and DALYs attributed to the selected risk factors were than estimated together with the PAF data from GBD 2001 study.The indirect method was employed to estimate the YLDs. Results: 51.09%of the total dearlls and 31.83%of the total DALYs from the Shandong population were resulted from the 19 selected risk factors.High blood pre.ure,smoking,low fruit and vegetable intake,aleohol consumption,indoor smoke from solid fuels,high cholesterol,urban air pollution, physical inactivity,overweight and obesity and unsafe injections in health care settings were identified as the top 10 risk faetors for mortality which together caused 50.21%of the total deaths.Alcohol use,smoking,high blood pressure,Low fruit and vegetable intake, indoor smoke from solid fuels, overweight and obesity,high cholesterol, physical inactivity,urban air pollution and iron-deficiency anemia were proved as the top 10 risk factors related to disease burden and were responsible for 29.04%of the total DALYs. Conclusion: Alcohol use.smoking and high blood pressure were determined as the major risk factors which influencing the health of residents in Shandong. The mortality and burden of disease could be reduced significantly if these major factors were effectively under control.

Molecular characterisation of environmental enterococci derived from water samples and assessment of associated public health hazards

Enterococci are versatile Gram-positive bacteria that can survive under extreme conditions. Most enterococci are non-virulent and found in the gastrointestinal tract of humans and animals. Other strains are opportunistic pathogens that contribute to a large number of nosocomial infections globally. Epidemiological studies demonstrated a direct relationship between the density of enterococci in surface waters and the risk of swimmer-associated gastroenteritis. The distribution of infectious enterococcal strains from the hospital environment or other sources to environmental water bodies through sewage discharge or other means, could increase the prevalence of these strains in the human population. Environmental water quality studies may benefit from focusing on a subset of Enterococcus spp. that are consistently associated with sources of faecal pollution such as domestic sewage, rather than testing for the entire genus. E. faecalis and E. faecium are potentially good focal species for such studies, as they have been consistently identified as the dominant Enterococcus spp. in human faeces and sewage. On the other hand enterococcal infections are predominantly caused by E. faecalis and E. faecium. The characterisation of E. faecalis and E. faecium is important in studying their population structures, particularly in environmental samples. In developing and implementing rapid, robust molecular genotyping techniques, it is possible to more accurately establish the relationship between human and environmental enterococci. Of particular importance, is to determine the distribution of high risk enterococcal clonal complexes, such as E. faecium clonal complex 17 and E. faecalis clonal complexes 2 and 9 in recreational waters. These clonal complexes are recognized as particularly pathogenic enterococcal genotypes that cause severe disease in humans globally. The Pimpama-Coomera watershed is located in South East Queensland, Australia and was investigated in this study mainly because it is used intensively for agriculture and recreational purposes and has a strong anthropogenic impact. The primary aim of this study was to develop novel, universally applicable, robust, rapid and cost effective genotyping methods which are likely to yield more definitive results for the routine monitoring of E. faecalis and E. faecium, particularly in environmental water sources. To fullfill this aim, new genotyping methods were developed based on the interrogation of highly informative single nucleotide polymorphisms (SNPs) located in housekeeping genes of both E. faecalis and E. faecium. SNP genotyping was successfully applied in field investigations of the Coomera watershed, South-East Queensland, Australia. E. faecalis and E. faecium isolates were grouped into 29 and 23 SNP profiles respectively. This study showed the high longitudinal diversity of E. faecalis and E. faecium over a period of two years, and both human-related and human-specific SNP profiles were identified. Furthermore, 4.25% of E. faecium strains isolated from water was found to correspond to the important clonal complex-17 (CC17). Strains that belong to CC17 cause the majority of hospital outbreaks and clinical infections globally. Of the six sampling sites of the Coomera River, Paradise Point had the highest number of human-related and human-specific E. faecalis and E. faecium SNP profiles. The secondary aim of this study was to determine the antibiotic-resistance profiles and virulence traits associated with environmental E. faecalis and E. faecium isolates compared to human pathogenic E. faecalis and E. faecium isolates. This was performed to predict the potential health risks associated with coming into contact with these strains in the Coomera watershed. In general, clinical isolates were found to be more resistant to all the antibiotics tested compared to water isolates and they harbored more virulence traits. Multi-drug resistance was more prevalent in clinical isolates (71.18% of E. faecalis and 70.3 % of E. faecium) compared to water isolates (only 5.66 % E. faecium). However, tetracycline, gentamicin, ciprofloxacin and ampicillin resistance was observed in water isolates. The virulence gene esp was the most prevalent virulence determinant observed in clinical isolates (67.79% of E. faecalis and 70.37 % of E. faecium), and this gene has been described as a human-specific marker used for microbial source tracking (MST). The presence of esp in water isolates (16.36% of E. faecalis and 19.14% of E. faecium) could be indicative of human faecal contamination in these waterways. Finally, in order to compare overall gene expression between environmental and clinical strains of E. faecalis, a comparative gene hybridization study was performed. The results of this investigation clearly demonstrated the up-regulation of genes associated with pathogenicity in E. faecalis isolated from water. The expression study was performed at physiological temperatures relative to ambient temperatures. The up-regulation of virulence genes demonstrates that environmental strains of E. faecalis can pose an increased health risk which can lead to serious disease, particularly if these strains belong to the virulent CC17 group. The genotyping techniques developed in this study not only provide a rapid, robust and highly discriminatory tool to characterize E. faecalis and E. faecium, but also enables the efficient identification of virulent enterococci that are distributed in environmental water sources.

Socioeconomic differences in takeaway food consumption among adults

Background In Australia and other developed countries, there are consistent and marked socioeconomic inequalities in health. Diet is a major contributing factor to the poorer health of lower socioeconomic groups: the dietary patterns of disadvantaged groups are least consistent with dietary recommendations for the prevention of diet-related chronic diseases compared with their more advantaged counterparts. Part of the reason that lower socioeconomic groups have poorer diets may be their consumption of takeaway foods. These foods typically have nutrient contents that fail to comply with the dietary recommendations for the prevention of chronic disease and associated risk factors. A high level of takeaway food consumption, therefore, may negatively influence overall dietary intakes and, consequently, lead to adverse health outcomes. Despite this, little attention has focused on the association between socioeconomic position (SEP) and takeaway food consumption, with the limited number of studies showing mixed results. Additionally, studies have been limited by only considering a narrow range of takeaway foods and not examining how different socioeconomic groups make choices that are more (or less) consistent with dietary recommendations. While a large number of earlier studies have consistently reported socioeconomically disadvantaged groups consume a lesser amount of fruit and vegetables, there is limited knowledge about the role of takeaway food in socioeconomic variations in fruit and vegetable intake. Furthermore, no known studies have investigated why there are socioeconomic differences in takeaway food consumption. The aims of this study are to: examine takeaway food consumption and the types of takeaway food consumed (healthy and less healthy) by different socioeconomic groups, to determine whether takeaway food consumption patterns explain socioeconomic variations in fruit and vegetable intake, and investigate the role of a range of psychosocial factors in explaining the association between SEP and takeaway food consumption and the choice of takeaway food. Methods This study used two cross-sectional population-based datasets: 1) the 1995 Australian National Nutrition Survey (NNS) which was conducted among a nationally representative sample of adults aged between 25.64 years (N = 7319, 61% response rate); and 2) the Food and Lifestyle Survey (FLS) which was conducted by the candidate and was undertaken among randomly selected adults aged between 25.64 years residing in Brisbane, Australia in 2009 (N = 903, 64% response rate). The FLS extended the NNS in several ways by describing current socioeconomic differences in takeaway food consumption patterns, formally assessing the mediated effect of takeaway food consumption to socioeconomic inequalities in fruit and vegetable intake, and also investigating whether (and which) psychosocial factors contributed to the observed socioeconomic variations in takeaway food consumption patterns. Results Approximately 32% of the NNS participants consumed takeaway food in the previous 24 hours and 38% of the FLS participants reported consuming takeaway food once a week or more. The results from analyses of the NNS and the FLS were somewhat mixed; however, disadvantaged groups were likely to consume a high level of �\less healthy. takeaway food compared with their more advantaged counterparts. The lower fruit and vegetable intake among lower socioeconomic groups was partly mediated by their high consumption of �\less healthy. takeaway food. Lower socioeconomic groups were more likely to have negative meal preparation behaviours and attitudes, and weaker health and nutrition-related beliefs and knowledge. Socioeconomic differences in takeaway food consumption were partly explained by meal preparation behaviours and attitudes, and these factors along with health and nutrition-related beliefs and knowledge appeared to contribute to the socioeconomic variations in choice of takeaway foods. Conclusion This thesis enhances our understanding of socioeconomic differences in dietary behaviours and the potential pathways by describing takeaway food consumption patterns by SEP, explaining the role of takeaway food consumption in socioeconomic inequalities in fruit and vegetable intake, and identifying the potential impact of psychosocial factors on socioeconomic differences in takeaway food consumption and the choice of takeaway food. Some important evidence is also provided for developing policies and effective intervention programs to improve the diet quality of the population, especially among lower socioeconomic groups. This thesis concludes with a discussion of a number of recommendations about future research and strategies to improve the dietary intake of the whole population, and especially among disadvantaged groups.

Implantation of osteogenic differentiated donor mesenchymal stem cells causes recruitment of host cells

The interaction between host and donor cells is believed to play an important role in osteogenesis. However, it is still unclear how donor osteogenic cells behave and interact with host cells in vivo. The purpose of this study was to track the interactions between transplanted osteogenic cells and host cells during osteogenesis. In vitro migration assay was carried out to investigate the ability of osteogenic differentiated humanmesenchymal stemcells (O-hMSCs) to recruit MSCs. At the in vivo level, O-hMSCs were implanted subcutaneously or into skull defects in severe combined immunodeficient (SCID) mice. New bone formation was observed bymicro-CT and histological procedures. In situ hybridization (ISH) against human Alu sequences was performed to distinguish donor osteogenic cells from host cells. In vitro migration assay revealed an increased migration potential of MSCs by co-culturing with O-hMSCs. In agreement with the results of in vitro studies, ISH against human Alu sequences showed that host mouse MSCs migrated in large numbers into the transplantation site in response to O-hMSCs. Interestingly, host cells recruited by O-hMSCs were the major cell populations in newly formed bone tissues, indicating that O-hMSCs can trigger and initiate osteogenesis when transplanted in orthotopic sites. The observations fromthis study demonstrated that in vitro induced O-hMSCs were able to attract hostMSCs in vivo andwere involved inosteogenesis togetherwith host cells,whichmay be of importance for bone tissue-engineering applications.

Experimental study on mechanical properties of pumpkin tissue

Purpose: The purpose of this study was to calculate mechanical properties of tough skinned vegetables as a part of Finite Element Modelling (FEM) and simulation of tissue damage during mechanical peeling of tough skinned vegetables. Design/methodology: There are some previous studies on mechanical properties of fruits and vegetables however, behaviour of tissue under different processing operations will be different. In this study indentation test was performed on Peel, Flesh and Unpeeled samples of pumpkin as a tough skinned vegetable. Additionally, the test performed in three different loading rates for peel: 1.25, 10, 20 mm/min and 20 mm/min for flesh and unpeeled samples respectively. The spherical end indenter with 8mm diameter used for the experimental tests. Samples prepare from defect free and ripped pumpkin purchased from local shops in Brisbane, Australia. Humidity and temperature were 20-55% and 20-250C respectively. Findings: Consequently, force deformation and stress and strain of samples were calculated and shown in presented figures. Relative contribution (%) of skin to different mechanical properties is computed and compared with data available from literature. According the results, peel samples had the highest value of rupture force (291N) and as well as highest value of firmness (1411Nm-1). Research limitations/implications: The proposed study focused on one type of tough skinned vegetables and one variety of pumpkin however, more tests will give better understandings of behaviours of tissue. Additionally, the behaviours of peel, unpeeled and flesh samples in different speed of loading will provide more details of tissue damages during mechanical loading. Originality/value: Mechanical properties of pumpkin tissue calculated using the results of indentation test, specifically the behaviours of peel, flesh and unpeeled samples were explored which is a new approach in Finite Element Modelling (FEM) of food processes. Keywords: Finite Element Modelling (FEM), relative contribution, firmness, toughness and rupture force.

Food consumption of Sri Lankan adults : an appraisal of serving characteristics

Objective The main aim of the present study was to identify food consumption in Sri Lankan adults based on serving characteristics. Design Cross-sectional study. Fruits, vegetables, starch, meat, pulses, dairy products and added sugars in the diet were assessed with portion sizes estimated using standard methods. Setting Twelve randomly selected clusters from the Sri Lanka Diabetes and Cardiovascular Study. Subjects Six hundred non-institutionalized adults. Results The daily intake of fruit (0·43), vegetable (1·73) and dairy (0·39) portions were well below national recommendations. Only 3·5 % of adults consumed the recommended 5 portions of fruits and vegetables/d; over a third of the population consumed no dairy products and fewer than 1 % of adults consumed 2 portions/d. In contrast, Sri Lankan adults consumed over 14 portions of starch and 3·5 portions of added sugars daily. Almost 70 % of those studied exceeded the upper limit of the recommendations for starch intake. The total daily number of meat and pulse portions was 2·78. Conclusions Dietary guidelines emphasize the importance of a balanced and varied diet; however, a substantial proportion of the Sri Lankan population studied failed to achieve such a recommendation. Nutrition-related diseases in the country may be closely correlated with unhealthy eating habits.

Diversity of dicotyledenous-infecting geminiviruses and their associated DNA molecules in Southern Africa, including the South-west Indian Ocean Islands

The family Geminiviridae comprises a group of plant-infecting circular ssDNA viruses that severely constrain agricultural production throughout the temperate regions of the world, and are a particularly serious threat to food security in sub-Saharan Africa. While geminiviruses exhibit considerable diversity in terms of their nucleotide sequences, genome structures, host ranges and insect vectors, the best characterised and economically most important of these viruses are those in the genus Begomovirus. Whereas begomoviruses are generally considered to be either monopartite (one ssDNA component) or bipartite (two circular ssDNA components called DNA-A and DNA-B), many apparently monopartite begomoviruses are associated with additional subviral ssDNA satellite components, called alpha- (DNA-αs) or betasatellites (DNA-βs). Additionally, subgenomic molecules, also known as defective interfering (DIs) DNAs that are usually derived from the parent helper virus through deletions of parts of its genome, are also associated with bipartite and monopartite begomoviruses. The past three decades have witnessed the emergence and diversification of various new begomoviral species and associated DI DNAs, in southern Africa, East Africa, and proximal Indian Ocean islands, which today threaten important vegetable and commercial crops such as, tobacco, cassava, tomato, sweet potato, and beans. This review aims to describe what is known about these viruses and their impacts on sustainable production in this sensitive region of the world. © 2012 by the authors licensee MDPI, Basel, Switzerland.

Food insecurity among adults residing in disadvantaged urban areas : potential health and dietary consequences

Objective: Food insecurity is the limited or uncertain availability or access to nutritionally-adequate, culturally-appropriate and safe foods. Food insecurity may result in inadequate dietary intakes, overweight or obesity and the development of chronic disease. Internationally, few studies have focused on the range of potential health outcomes related to food insecurity among adults residing in disadvantaged locations and no such Australian studies exist. The objective of this study was to investigate associations between food insecurity, socio-demographic and health factors and dietary intakes among adults residing in disadvantaged urban areas. Design: Data were collected by mail survey (n= 505, 53% response rate), which ascertained information about food security status, demographic characteristics (such as age, gender, household income, education) fruit and vegetable intakes, take-away and meat consumption, general health, depression and chronic disease. Setting: Disadvantaged suburbs of Brisbane city, Australia, 2009. Subjects: Individuals aged ≥ 20 years. Results: Approximately one-in-four households (25%) were food insecure. Food insecurity was associated with lower household income, poorer general health, increased healthcare utilisation and depression. These associations remained after adjustment for age, gender and household income. Conclusion: Food insecurity is prevalent in urbanised disadvantaged areas in developed countries such as Australia. Low-income households are at high risk of experiencing food insecurity. Food insecurity may result in significant health burdens among the population, and this may be concentrated in socioeconomically-disadvantaged suburbs.

Potential dietary consquences of food insecurity among Australian adults residing in urbanised disadvantaged areas

Introduction: Food insecurity is the limited/uncertain availability, access to or ability to acquire nutritionally-adequate, culturallyrelevant and safe foods. Adults suffering from food insecurity are at risk of inadequate nutrient intakes or, paradoxically, overweight/ obesity and the development of chronic disease. Despite the global financial crisis and rising costs of living, there are few studies investigating the potential dietary consequences of food insecurity among the Australian population. This study examined whether food insecurity was associated with weight status and poorer intakes of fruits, vegetable and takeaway foods among adults residing in socioeconomically-disadvantaged urbanised areas. Methods: In this cross-sectional study, a random sample of residents (n=1000) were selected from the most disadvantaged suburbs of Brisbane city (response rate 51%). Data were collected by postal questionnaire which ascertained information on sociodemographic information, household food security status, height, weight, fruit and vegetable intakes and takeaway consumption. Data were analysed using chi-square and logistic regression. Results: The overall prevalence of food insecurity was 31%. Food insecurity was not associated with weight status among men or women. Associations between food security status and potential dietary consequences differed for men and women. Among women, food security was not associated with intakes of fruit, vegetable or takeaway consumption. Contrastingly, among men food security was associated with vegetable intakes and consumption of takeaway food: men reporting food insecurity had lower intakes of vegetables and were more likely to consume takeaway foods compared to those that were food secure. Conclusion: Food security is an important public health issue in Australia and has potential dietary consequences that may adversely affect the health of food-insecure groups, most notably men residing in food-insecure households.

What are the implications of peri-urban agriculture on food security in Australian cities?

Background: Periurban agriculture refers to agricultural practice occurring in areas with mixed rural and urban features. It is responsible 25% of the total gross value of economic production in Australia, despite only comprising 3% of the land used for agriculture. As populations grows and cities expand, they are constantly absorbing surrounding fringe areas, thus creating a new fringe, further from the city causing the periurban region to constantly shift outwards. Periurban regions are fundamental in the provision of fresh food to city populations and residential (and industrial) expansion taking over agricultural land has been noted as a major worldwide concern. Another major concern around the increase in urbanisation and resultant decrease in periurban agriculture is its potential effect on food security. Food security is the availability or access to nutritionally-adequate, culturally-relevant and safe foods in culturally-appropriate ways. Thus food insecurity occurs when access to or availability of these foods is compromised. There is an important level of connectedness between food security and food production and a decrease in periurban agriculture may have adverse effects on food security. A decrease in local, seasonal produce may result in a decrease in the availability of products and an increase in cost, as food must travel greater distances, incurring extra costs present at the consumer level. Currently, few Australian studies exist examining the change in periurban agriculture over time. Such information may prove useful for future health policy and interventions as well as infrastructure planning. The aim of this study is to investigate changes in periurban agriculture among capital cities of Australia. Methods: We compared data pertaining to selected commodities from the Australian Bureau of Statistics 2000-01 and 2005 -2006 Agricultural Census. This survey is distributed online or via mail on a five-yearly basis to approximately 175,000 Agricultural business to ascertain information on a range of factors, such as types of crops, livestock and land preparation practices. For the purpose of this study we compared the land being used for total crops, and cereal , oil seed, legume, fruit and vegetable crops separately. Data was analysed using repeated measures anova in spss. Results: Overall, total area available for crops in urbanised areas of Australia increased slightly by 1.8%. However, Sydney, Melbourne, Adelaide and Perth experienced decreases in the area available for fruit crops by 11%, 5%,and 4% respectively. Furthermore, Brisbane and Perth experienced decreases in land available for vegetable crops by 28% and 14% respectively. Finally, Sydney, Adelaide and Perth experienced decreases in land available for cereal crops by 10 – 79%. Conclusions: These findings suggest that population increases and consequent urban sprawl may be resulting in a decrease in peri-urban agriculture, specifically for several core food groups including fruit, breads and grain based foods. In doing so, access to or availability of these foods may be limited, and the cost of these foods is likely to increase, which may compromise food insecurity for certain sub-groups of the population.

Comparing the availability, price, variety and quality of fruits and vegetables across retail outlets and by area level socioeconomic position

Abstract Objective: To explore whether area-level socioeconomic position or the form of retail stream (conventional versus farmers’ market) are associated with differences in the price, availability, variety and quality of a range of fresh fruit and vegetables. Design: A multi-site cross-sectional pilot study of farmers’ markets, supermarkets and independent fruit and vegetable retailers. Each was surveyed to assess the price, availability, variety and quality of 15 fruit and 18 vegetable items. Setting: Retail outlets were located in South-East Queensland. Subjects: Fifteen retail outlets were surveyed (five of each retail stream). Results: Average basket prices were not significantly different across the socioeconomic spectrum however prices in low socioeconomic areas were cheapest. Availability, variety, and quality did not differ across levels of socioeconomic position however the areas with the most socioeconomic disadvantage scored poorest for quality and variety. Supermarkets had significantly better fruit and vegetable availability than farmers’ markets however price, variety and quality scores were not different across retail streams. Results demonstrate a trend to fruit and vegetable prices being more expensive at farmers’ markets, with the price of the Fruit basket being significantly greater at the organic farmer’s market compared with the non-organic farmers’ markets. Conclusions: Neither area-level socioeconomic position nor the form of retail stream was significantly associated with differences in the availability, price, variety and quality of fruit and vegetables, except for availability which was higher in supermarkets than farmers’ markets. Further research is needed to determine what role farmers’ markets can play in affecting fruit and vegetable intake.

Cytokine synthesis by intestinal intraepithelial lymphocytes. Both gamma/delta T cell receptor-positive and alpha/beta T cell receptor-positive T cells in the G1 phase of cell cycle produce IFN-gamma and IL-5

Murine intestinal intraepithelial lymphocytes (IEL) have been shown to contain subsets of alpha/beta TCR+ and gamma/delta TCR+ T cells that spontaneously produce cytokines such as IFN-gamma and IL-5. We have now determined the nature and cell cycle stage of these cytokine-producing T lymphocytes in EIL by using IFN-gamma- and IL-5-specific ELISPOT assay, cytokine-specific mRNA-cDNA dot-blot hybridization and polymerase chain reaction, and flow cytometry (FACS) for DNA analysis. When CD3+ T cells from IEL of normal C3H/HeN mice were separated into low and high density fractions by discontinuous Percoll gradients, IFN-gamma and IL-5 spot-forming cells were only found in the former population. Analysis of mRNA for these cytokines by both IFN-gamma- and IL-5-specific dot-blot hybridization and polymerase chain reaction revealed that higher levels of message for IFN-gamma and IL-5 were also seen in the low density fraction. However, cell cycle analysis of these two fractions by FACS using propidium iodide showed a similar pattern of cell cycle stages in both low and high density populations (G0 + G1 approximately 96 to 98% and S/G2 + M approximately 2 to 4%). Finally, mRNA from gamma/delta TCR+ and alpha/beta TCR+ T cells in both low and high density fractions of IEL were analyzed for IFN-gamma and IL-5 message by polymerase chain reaction. After 35 cycles of amplification, both gamma/delta TCR+ and alpha/beta TCR+ T cells in the low density fraction expressed higher levels of message for these two cytokines when compared with the high density population. These results have now shown that both gamma/delta and alpha/beta TCR+ IEL can be separated into low and high density subsets and both fractions possess a similar stage of cell cycle. However, only the low density cells (in G1 phase) of both gamma/delta and alpha/beta TCR types possess increased cytokine-specific mRNA and produce the cytokines IFN-gamma and IL-5. Our results suggest that alpha/beta TCR+ and gamma/delta TCR+ IEL can produce cytokines without cell proliferation.