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Cell culture contamination: sources, consequences, prevention, and elimination.

The subject of the chapter is cell culture contamination. Contamination may enter the cell culture system as a physical, chemical, and/or biological component of the environment. The potential sources and consequences of cell culture contamination are unique to the cell culture system and the contaminant. A basic understanding of cell culture contamination is necessary to appreciate the need to develop and practice standardized cell culture procedures. General sources, consequences, and preventative measures are discussed for physical and chemical contamination based on current technology. Mycoplasmal contamination is the focus of the discussion on biological contamination and its impact on cell cultures. The introduction of other biological contaminants should be controlled by the institution of cell culture management procedures needed to minimize the incidence of mycoplasmal contamination. The need to eliminate the routine use of antibiotics in cell culture systems and institute routine testing to detect contamination is emphasized. More rapid detection of contamination should reduce the incidence of cross-contamination and minimize the consequences of any contamination event.

Animals↗

The neurobiological consequences of early stress and childhood maltreatment.

Early severe stress and maltreatment produces a cascade of neurobiological events that have the potential to cause enduring changes in brain development. These changes occur on multiple levels, from neurohumoral (especially the hypothalamic-pituitary-adrenal [HPA] axis) to structural and functional. The major structural consequences of early stress include reduced size of the mid-portions of the corpus callosum and attenuated development of the left neocortex, hippocampus, and amygdala. Major functional consequences include increased electrical irritability in limbic structures and reduced functional activity of the cerebellar vermis. There are also gender differences in vulnerability and functional consequences. The neurobiological sequelae of early stress and maltreatment may play a significant role in the emergence of psychiatric disorders during development.

Adolescent↗

Low doses of corticotropin-releasing hormone injected into the dorsal raphe nucleus block the behavioral consequences of uncontrollable stress.

The behavioral consequences of uncontrollable stress that are collectively called learned helplessness (LH) are mediated in part by increased levels of serotonin (5-HT) activity in the dorsal raphe nucleus (DRN) and it's projection regions. Recently, corticotropin-releasing hormone (CRH) within the DRN has been implicated in the development of LH because intra-DRN CRH produces LH at very high doses, and because intra-DRN antagonists for the CRH 2 receptor (CRHR2) block LH. Since these behavioral effects are mediated by both 5-HT excitation and CRHR2 activation, we have suggested that CRHR2 mediates excitation of DRN 5-HT neurons. However, CRH has been shown to inhibit DRN 5-HT neurons at low doses that are expected to bind to CRHR1. Since CRHR1 antagonists were ineffective in blocking LH, we have further suggested that CRHR1 might mediate the inhibition of DRN 5-HT neurons. In support of this hypothesis, although low doses of CRH that preferentially bind CRHR1 inhibit DRN 5-HT activity, higher doses at which CRH would be expected to bind both receptor subtypes no longer inhibit DRN 5-HT. In addition, high doses of CRH are required to produce LH, which is known to be mediated by 5-HT excitation, and the CRHR2 agonist urocortin II (UCN II) produces LH at much lower doses than does CRH. The present studies show that intra-DRN CRH microinjection blocks the behavioral effects produced by DRN UCN II, but only at doses that have been shown to inhibit DRN 5-HT activity. Indeed, a higher dose of CRH that has been shown to no longer inhibit DRN 5-HT activity did not affect the behavioral consequences of DRN UCN II. In a separate experiment, the effective dose of CRH blocked the usual behavioral consequences of uncontrollable stress.

Animals↗

Acute and long-term consequences of single MDMA administration in relation to individual anxiety levels in the rat.

Our previous work has shown that normal male Wistar rats can differ systematically in their behavioral response to the elevated plus-maze (EPM), where animals with high (HA) or low anxiety (LA) levels can be identified based on the percentage of time spent in the open arms. These animals also differ in other behavioral tests (e.g. active avoidance), and in their serotonin levels in the ventral striatum. Here, we tested whether such HA and LA rats might respond differently to the amphetamine analogue 3,4-methylenedioxymethamphetamine (MDMA, "ecstasy"). This drug can affect psychomotor activation and anxiety; effects which are probably due to its pronounced serotonergic and dopaminergic impacts in the rat brain. Based on a routine screening procedure in the plus-maze, male Wistar rats were divided into HA and LA sub-groups, in which rectal temperature was measured. Thirty minutes after the i.p. injection of MDMA (7.5 or 15 mg/kg) or vehicle, they were again tested in the plus-maze. During the next 3 weeks, the animals underwent further behavioral tests (plus-maze, open field, active avoidance, forced swimming) to test for possible long-term consequences of MDMA. Rectal temperature was found to be higher in LA than HA rats and was especially increased with the higher dose of MDMA (15 mg/kg). In the acute plus-maze test, the lower dose of MDMA led to an anxiogenic-like profile, whereas the higher dose led to an anxiolytic-like profile, both in HA and LA rats. Possible long-term consequences of MDMA were only tested with 7.5 mg/kg MDMA, since the 15 mg/kg dose led to a high level of lethality. The analysis of open field, plus-maze (performed after 9-12 days), and forced swimming behavior (performed after 20-21 days) did not provide indications for lasting effects of MDMA. In contrast, active avoidance learning was impaired in LA- but not HA-rats treated with MDMA. A single injection of MDMA does not only have acute effects on anxiety and psychomotor activation, but can also have some prolonged or delayed task-dependent behavioral consequences. The detection of such sequels can require that individual differences are taken into account and here, determining anxiety levels in the EPM seems to serve as a useful approach.

Analysis of Variance↗

A model to estimate the financial consequences of classical swine fever outbreaks: principles and outcomes.

A model is presented aimed at a financial analysis of a Classical Swine Fever outbreak. Financial consequences are calculated for affected parties, including governments (EU and national), farms, and related industries in the production chain. The model can be used to calculate the losses of a real outbreak as well as of a simulated one. In this article, the model is applied to the 1997/1998 outbreak of Classical Swine Fever in the Netherlands. Results show that total financial consequences of the outbreak are US $2.3 billion. Consequential losses for farmers and related industries are US $423 million and US $596 million respectively. Budgetary consequences for governments include less than 50% of the total losses calculated by the model. The model can be adapted easily to suit other diseases and countries.

Animal Husbandry↗

Waiting times for radiotherapy: consequences of volume increase for the TCP in oropharyngeal carcinoma.

BACKGROUND AND PURPOSE: Waiting lists for radiotherapy have become longer over the past years. Apart from the psychological distress for the patient we are concerned about tumour growth during this waiting time, which may worsen prognosis. The purpose of this pilot study was to investigate tumour growth in the waiting time and to obtain an indication of its clinical consequences for patients with oropharyngeal carcinoma. A tumour control probability (TCP) model was applied to evaluate consequences for outcome. METHODS AND MATERIALS: Increase in tumour volume was measured for 13 patients with oropharyngeal carcinoma by outlining the tumour on the diagnostic as well as on the treatment planning CT scan. Waiting time was defined as time between histopathological diagnosis and start of radiotherapy. For each tumour we calculated the increase in tumour volume and the tumour doubling time. The potential increase in TCP was calculated for each tumour for the situation without treatment delay. RESULTS: The mean increase in tumour volume was 70%. The mean waiting time was 56 days. Expected TCP with incorporation of delay was 47%, without delay it might have been 63-66%. CONCLUSION: This study shows tumour progression during the time between the diagnostic CT scan and the treatment planning CT scan in oropharyngeal cancer. As a consequence of waiting time, which allows tumour volume increase, there may be an average control loss of 16-19 % for these tumours during the total waiting time before radiotherapy.

Aged↗

[Patients admitted to the intensive care unit for iatrogenic disease. Risk factors and consequences].

PURPOSE: Data pertaining to iatrogenic diseases have been recorded over the last 25 years. Regarding the evolution of medical practice (general ageing, more and more powerful drugs and complex procedures), it is not known whether the incidence and the consequences of iatrogenic diseases have changed since their first evaluation. METHODS: To determine the admission rate to intensive care units for iatrogenic diseases, with the purpose of analyzing risk factors and consequences, and to compare our results with previous data recorded in 1979 (admission rate: 12.6%, mortality: 20%, preventable events: 47%), a 1-year retrospective study was conducted in an intensive care unit (ICU). RESULTS: During 1994, 68 (10.9%) out of 623 patients were admitted to the ICU for iatrogenic diseases (drugs: 41, medical acts: 12, surgical acts: 15). They were not different--in terms of severity, mortality, workload and length of stay in the ICU--from the other 555 patients hospitalized for other reasons. They were hospitalized on average for 472 days in the ICU, with a 13% fatality rate and a financial cost of US $688,470. Risk factors for iatrogenic diseases were the age and the number of prescribed drugs. The rate of preventable events was 51%. CONCLUSIONS: In this study, the occurrence of life-threatening iatrogenic diseases was a persistent and important purpose for admission to the ICU. Risk factors and consequences are still identical to those reported in 1979. Our results emphasize the persistence of the noxious impact of iatrogenic diseases on the quality and cost of medical care.

Adult↗

Human schistosomiasis: potential long-term consequences of vaccination programmes.

Fields trials of new schistosomiasis vaccines are anticipated within the next few years, but there remains great uncertainty regarding the optimal design of vaccination programmes. Mathematical models are used here to explore the potential long-term consequences of vaccination, assuming that the vaccines provide partial protection for a limited period. The analysis suggests that vaccines acting to reduce infection rates or egg output will have a similar impact on levels of infection, that this impact may be highly sensitive to the duration as well as the degree of protection, that it may take several decades for the full impact to become apparent, and that one consequence will be peak levels of infection occurring in older age classes. In terms of lowering levels of infection there may be advantages in delaying vaccination until children reach school age, especially if the vaccine gives short-lived protection, or to repeat vaccination. The short-term advantages can be greatly increased by combining the introduction of a vaccination programme with initial mass chemotherapy. Continuous combined vaccination and chemotherapy programmes may also be more effective than either intervention alone. More research is needed on the consequences of vaccinating previously vaccinated, infected, and infected and treated individuals and the importance of natural boosting of vaccine-induced immunity.

Adolescent↗

A model for evaluating physico-chemical substance properties required by consequence analysis models.

Modeling systems for analyzing the consequences of chemical emergencies require as input values a number of physico-chemical substance properties, commonly as a function of temperature at atmospheric pressure. This paper presents a mathematical model "CHEMIC", which can be used for evaluating such substance properties, assuming that six basic constant quantities are available (molecular weight, freezing or melting point, normal boiling point, critical temperature, critical pressure and critical volume). The model has been designed to yield reasonably accurate numerical predictions, while at the same time keeping the amount of input data to a minimum. The model is based on molecular theory or thermodynamics, together with empirical corrections. Mostly, model equations are based on the so-called law of corresponding states. The model evaluates substance properties as a function of temperature at atmospheric pressure. These include seven properties commonly required by consequence analysis and heavy gas dispersion modeling systems: vapor pressure, vapor and liquid densities, heat of vaporization, vapor and liquid viscosities and binary diffusion coefficient. The model predictions for vapor pressure, vapor and liquid densities and heat of vaporization have been evaluated by using the Clausius-Clapeyron equation. We have also compared the predictions of the CHEMIC model with those of the DATABANK database (developed by the AEA Technology, UK), which includes detailed semi-empirical correlations. The computer program CHEMIC could be easily introduced into consequence analysis modeling systems in order to extend their performance to address a wider selection of substances.

Atmospheric Pressure↗

Acute public health consequences associated with hazardous substances released during transit, 1993-2000.

Massive quantities of hazardous substances are transported each day throughout the United States. While most arrive safely at their destination, uncontrolled releases of substances in transit do occur and have the potential of causing acute public health consequences for those individuals at or near the release. Data from 16 state health departments participating in the Agency for Toxic Substances and Disease Registry's (ATSDR) Hazardous Substances Emergency Events Surveillance (HSEES) system were analyzed to determine the public health consequences that occurred from actual releases in transit. Of the 9392 transportation events analyzed, 9.1% resulted in 2008 victims, including 115 deaths. The population groups injured most often were employees and the general public. The most common injury sustained was respiratory irritation. Evacuations were ordered in 5.5% of events effecting at least 63,686 people. Human error and equipment failure were the most common factors leading to events. These findings underscore the importance of job safety training, community planning, and effective emergency response to prevent adverse public health consequences from occurring or lessen their effect on the public.

Disasters↗

Uncertainty quantification in the health consequences of the boiling liquid expanding vapour explosion phenomenon.

A methodology for estimating the risk owing to the phenomenon of boiling liquid expanding vapour explosion (BLEVE) in the presence of uncertainties both in the model and in the parameters of the models is presented. BLEVE takes place when a tank containing liquefied petroleum gas (LPG) is exposed to fire and fails catastrophically. Two models have been used in the estimation of the intensity of thermal radiation from the resulting fireball, namely the solid-flame model assuming an emission power independent of the combustion mass and the point-source model that estimates the emissive power as a function of the combustion mass. Three measures of the BLEVE consequences, the intensity of thermal radiation, the dose of thermal radiation and the probability of loss of life as a result of the exposure to the thermal radiation and as a function of the distance from the center of the tank have been considered. Uncertainties in the exact values of the parameters of the models have been quantified and the resulting uncertainties in the three consequence measures have been assessed. A sensitivity analysis on the relative contribution of the uncertainty in each of the input variables to the uncertainties of the consequence measures has been performed. One conclusion is that the uncertainties in the probability of loss of life are mainly due to the uncertainties in the model of the physical phenomenon rather than to the uncertainties of the dose-response model.

Accidents, Occupational↗

Musculoskeletal pain in the Netherlands: prevalences, consequences and risk groups, the DMC(3)-study.

The objective of this paper was to present estimates on the prevalence of musculoskeletal pain of five different anatomical areas and ten anatomical sites, and their consequences and risk groups in the general Dutch population. Cross-sectional data from a population-based study of a sex-age stratified sample of Dutch inhabitants of 25 years and older were used. With a postal questionnaire data was assessed on musculoskeletal pain, additional pain characteristics (location, duration, course), its consequences (utilization of health care, sick leave and limitation in daily life) and general socio-demographic characteristics. The top three of self-reported musculoskeletal pain (point prevalence (P(p)) with 95% confidence interval (CI)) was: (1). low back pain, P(p)=26.9% (95% CI 25.5-28.3); (2). shoulder pain, P(p)=20.9% (95% CI 19.6-22.2); and (3). neck pain, P(p)=20.6% (95% CI 19.3-21.9). In most cases the pain was described as continuous or recurrent and mild. In every three out of ten cases the complaints about pain were accompanied by limitations in daily living. Between 33 and 42% of those with complaints consulted their general practitioner about their pain. With the exception of persons who are work disabled, general sociodemographic characteristics cannot be used to identify high risk groups. Musculoskeletal pain is common in all subgroups of the population and has far-reaching consequences for health, work and the use of health care.

Adult↗

Consequences of repeated early isolation in domestic piglets (Sus scrofa) on their behavioural, neuroendocrine, and immunological responses.

Stress in the form of intermittent maternal deprivation and social isolation during early postnatal life in rats and monkeys produces persistent changes in physiology and behaviour. In farm animals physiological consequences of disrupting mother-infant interactions with respect to health and animal welfare are relatively unknown. Therefore, the aim of the present study was to investigate the behavioural, neuroendocrine and immunological consequences of a 2 h daily social isolation from day 3 to day 11 of age in domestic piglets as well as potential long-term effects on the brain-endocrine-immune regulation. Repeated social isolation resulted in significantly decreased open-field activity (locomotion, vocalization) during the isolation period, increased basal cortisol concentrations and decreased lymphocyte proliferation in response to concanavalin A and pokeweed mitogen one day after the isolation. There was also a significant increase of interleukin-1beta (IL-1beta) concentration in hippocampus in isolated piglets compared to controls at this time. Six weeks after isolation significant enhanced basal ACTH concentrations as well as higher IL-1beta content and glucocorticoid receptor (GR) binding in hippocampus were found. These endocrine and immune responses were associated with decreased CRH levels in the hypothalamus and increased CRH content in the amygdala. The present data indicate that early social isolation in pigs may cause changes in behavioural, neuroendocrine, and immune regulation and produce long-term effects not only on the activity of the hypothalamic-pituitary-adrenal (HPA) system, but also on the immune-brain circuitry with possible negative consequences in health and welfare of commercial pigs. Using the pig as a suitable animal model, the finding of this study may also have some implications for the etiology of anxiety and depression in humans.

Adrenocorticotropic Hormone↗

Prevalence and consequences of asthma and wheezing in African-American and White adolescents.

PURPOSE: To describe the functional consequences of health care utilization for wheezing symptoms and diagnosed asthma in a multiracial school-based population. METHODS: Wheezing symptoms and physician-diagnosed asthma were measured in 2059 eighth graders with the International Study of Asthma and Allergies in Children video questionnaire. The sample site population was 40% African-American, 53% White, and 49% girls. Thirty-two percent of the children were on the free school lunch program. Adolescents self-reported their wheezing symptoms and physician diagnosis in response to the video wheezing scenes. They also answered questions on activity limitations, school attendance, and sleep disturbances owing to their wheezing symptoms. Multiple logistic regression was used to calculate odds ratios and adjust for potential confounders. RESULTS: Nine percent of the adolescents reported physician-diagnosed asthma with current symptoms and 27% reported current wheezing with no diagnosis of asthma. Physician-diagnosed asthmatics and wheezers were 2.6 (95% confidence interval [CI] 1.9, 3.6) and 1.8 (95% CI 1.4, 2.2) times more likely, respectively, to miss school days because of wheezing symptoms than asymptomatic adolescents. Diagnosed asthmatics were 7.8 (95% CI 5.5, 11.2) times and wheezers 4.7 (95% CI 3.5, 6.1) times more likely to have sleep disturbances than asymptomatic adolescents. Diagnosed asthmatics were 49 (95% CI 30.0, 79.8) times and wheezers 4.8 (95% CI 3.0, 7.5) times as likely to report a clinic visit for wheezing than asymptomatic adolescents. African-Americans had similar prevalence of wheezing, compared with Whites, although African-Americans were 1.4 (95% CI 1.1, 1.9) times as likely to have physician-diagnosed asthma. Girls were twice as likely to report physician-diagnosed asthma and wheezing symptoms as were boys. CONCLUSIONS: Adolescents with wheezing experienced functional consequences comparable to those of adolescents with physician-diagnosed asthma. Although wheezers may have less severe asthma than diagnosed asthmatics, the functional consequences of wheezing are likely to impair school performance and limit activity.

Black or African American↗

Consequences of feeding for future feeding.

The intake of food has physiological consequences via physical (e.g. distension) and chemical (e.g. glucose) stimulation of receptors in the viscera and, in the longer term, by changes in signals from adipose tissue (e.g. leptin), integrated by the CNS. These consequences are associated with the sensory properties of the food such that repeated exposure to a food generates a conditioned acceptance or rejection reflex with the physiological consequences of eating as the unconditioned stimulus (US) and the sensory characteristics of the food as the conditioned stimulus (CS). Such learnt preferences and aversions occur throughout the animal kingdom, from nematodes to human beings, with much of the research being carried out with insects, laboratory animals and farm animals. Preferences for and aversions to particular foods are manifested in non-random choices between two or more foods on offer but also influence the quantity eaten when only one food is available. These considerations have been developed into a theory of Minimal Total Discomfort which proposes that an animal experiments with the amount eaten per day, and its selection between different foods, until the total of the signals generated from excesses or deficiencies of food components is minimised. Changes in food composition and/or nutrient requirements can therefore be matched by appropriate changes in intake and selection.

Animals↗

Malaria in adolescence: burden of disease, consequences, and opportunities for intervention.

The problem of malaria in adolescence has largely been overshadowed by the huge burden of disease in young children. A substantial number of adolescents are at risk from malaria infection, but the burden of disease and consequences of infection in this age-group have rarely been studied. Our understanding of specific risk factors and beneficial interventions for adolescents is also limited. Data show that, from an adolescent viewpoint, malaria is a common cause of clinical illness and a preventable cause of death, even in areas of stable malaria transmission. Younger adolescents might be at a higher risk than older adolescents, because of immunological and hormonal factors. There are limited data about the adverse consequences of malaria in non-pregnant adolescents. However, in pregnant adolescents, the consequences of malaria are of great concern and simple interventions might lead to a substantial benefit. Malaria infection in adolescents is an under-recognised problem, and the prevention, diagnosis, and treatment of malaria should have a high priority within adolescent health programmes.

Adolescent↗

Psychological consequences of road traffic accidents for children and their mothers.

BACKGROUND: Little is known about the psychological and behavioural consequences of road traffic accidents for children. The study aimed to determine the outcome of road traffic accidents on children and their mothers. METHOD: A 1-year cohort study of consecutive child attenders aged 5-16 years at an Accident and Emergency Department. Data were extracted from medical notes and from interview and self-report at baseline, 3 months and 6 months. RESULTS: The children had an excellent physical outcome. Fifteen per cent suffered acute stress disorder; 25% suffered post-traumatic stress disorder at 3 months and 18% at 6 months. Travel anxiety was frequent. Post-traumatic consequences for mothers were common. CONCLUSION: Psychological outcome was poor for a minority of children and associated with disability, especially for travel. There were significant family consequences. There is a need for changes in clinical care to prevent, identify and treat distressing and disabling problems.

Accidents, Traffic↗

Consequences of major and minor depression in later life: a study of disability, well-being and service utilization.

BACKGROUND: The consequences of major depression for disability, impaired well-being and service utilization have been studied primarily in younger adults. In all age groups the consequences of minor depression are virtually unknown. In later life, the increased co-morbidity with physical illness may modify the consequences of depression, warranting special study of the elderly. With rising numbers of elderly people, excess service utilization by depressed elderly represents an increasingly important issue. METHODS: Based on a large, random community-based sample of older inhabitants of the Netherlands (55-85 years), the associations of major and minor depression with various indicators of disability, well-being and service utilization were assessed, controlling for potential confounding factors. Depression was diagnosed using a two-stage screening design. Diagnosis took place in all subjects with high depressive symptom levels and a random sample of those with low depressive symptom levels. The study sample consists of all participants to diagnostic interviews (N = 646). RESULTS: As in younger adults, associations of both major and minor depression with disability and well-being remained significant after controlling for chronic disease and functional limitations. Adequate treatment is often not administered, even in subjects with major depression. As the vast majority of those depressed were recently seen by their general practitioners, treatment could have been provided in most cases. Bivariate analyses show that major and minor depression are associated with an excess use of non-mental health services, underscoring the importance of recognition. In multivariate analyses the evidence of excess service utilization was less compelling. CONCLUSIONS: Both major and minor depression are consequential for well-being and disability, supporting efforts to improve the recognition and treatment in primary care. However, controlled trials are necessary to assess the impact this may have on service utilization.

Activities of Daily Living↗