Quantitative risk assessment for developmental toxicity.

Pharmaceutical companies and governmental regulatory agencies are becoming increasingly aware of the need for improved statistical methods for developmental toxicity experiments. Although a number of statisticians have become interested in this area, activity has centered mostly on the development of methods to analyze binary outcomes, such as malformations among live pups, while accounting appropriately for the correlation induced by the litter effect. In contrast, the topic of quantitative risk assessment has received relatively little attention. This paper addresses the specific question of how to assess risk appropriately when exposure causes a variety of adverse effects, including resorption and fetal death, in addition to malformations. It will be seen that risk assessments based on a single developmental outcome, such as malformation, may be conservative. A method is proposed for estimating an exposure level at which the overall risk of any adverse effect is acceptably low. The method is based on a continuation ratio formulation of a multinomial distribution, with an additional scale parameter to account for overdispersion. Comparisons are made with binary models on prenatal death and malformation, as well as a binary model that makes no distinction between death and malformation, but simply classifies each fetus as normal or abnormal. Data from several developmental toxicity studies illustrate the results and findings.     

Quantitative microbiological risk assessment: principles applied to determining the comparative risk of salmonellosis from chicken products

Since causal links have been established between food-borne illness and particular microorganisms, it has been possible to assess the public health risks of their presence in foods and propose measures to ensure the safety of customers. Effective control measures for food safety have been based on knowledge of the resistance of pathogenic microorganisms to the treatments used for preservation (e.g. acidification or reduced water activity) or decontamination (e.g. pasteurisation or sterilisation). Using this principle, food safety has been managed informally and successfully within the food industry for many years. Recently, formal risk assessment schemes, for example from Codex Alimentarius, have developed and placed the elements of decision-making on suitable control measures into a formal framework with clearly identifiable stages. The output of the four stages of a formal risk assessment (hazard identification, hazard characterisation, exposure assessment and risk characterisation) provides the basis for decisions on actions needed to control the identified hazard. There are many difficulties in ensuring that risk assessments are realistic and accessible to potential users, in many cases their value is limited by the data available. The study reported here on control of Salmonella in poultry products illustrates that it is possible to produce a comparative risk assessment based on published data. This study is not a full quantitative risk assessment, but provide useful pointers for a risk manager. Differences are discussed between the exposure assessment data needed to propose controls for infectious or toxigenic pathogens. For infectious pathogens, the presence of viable and infectious microorganisms is itself the hazard, but for toxigenic microorganisms, absence or destruction of viable cells at ingestion does not in itself ensure the absence of toxin. For toxin hazards, exposure assessment needs to consider previous conditions that may have led to toxin formation and persistence, rather than just the level of microbes at ingestion.     

Quantitative microbial risk assessment models for consumption of raw vegetables irrigated with reclaimed water

Quantitative microbial risk assessment models for estimating the annual risk of enteric virus infection associated with consuming raw vegetables that have been overhead irrigated with nondisinfected secondary treated reclaimed water were constructed. We ran models for several different scenarios of crop type, viral concentration in effluent, and time since last irrigation event. The mean annual risk of infection was always less for cucumber than for broccoli, cabbage, or lettuce. Across the various crops, effluent qualities, and viral decay rates considered, the annual risk of infection ranged from 10(-3) to 10(-1) when reclaimed-water irrigation ceased 1 day before harvest and from 10(-9) to 10(-3) when it ceased 2 weeks before harvest. Two previously published decay coefficients were used to describe the die-off of viruses in the environment. For all combinations of crop type and effluent quality, application of the more aggressive decay coefficient led to annual risks of infection that satisfied the commonly propounded benchmark of < or =10(-4), i.e., one infection or less per 10,000 people per year, providing that 14 days had elapsed since irrigation with reclaimed water. Conversely, this benchmark was not attained for any combination of crop and water quality when this withholding period was 1 day. The lower decay rate conferred markedly less protection, with broccoli and cucumber being the only crops satisfying the 10(-4) standard for all water qualities after a 14-day withholding period. Sensitivity analyses on the models revealed that in nearly all cases, variation in the amount of produce consumed had the most significant effect on the total uncertainty surrounding the estimate of annual infection risk. The models presented cover what would generally be considered to be worst-case scenarios: overhead irrigation and consumption of vegetables raw. Practices such as subsurface, furrow, or drip irrigation and postharvest washing/disinfection and food preparation could substantially lower risks and need to be considered in future models, particularly for developed nations where these extra risk reduction measures are more common.     

纳米银胶/壳聚糖复合抗菌剂的制备及其在洗涤产品中抗菌性能

本文研究了纳米银胶/壳聚糖抗菌剂的制备及其形貌的表征分析,以大肠杆菌为代表菌株,研究了复合抗菌剂在洗涤产品中的抗菌效率及抗菌的稳定性,结果说明复合抗菌剂在洗涤产品中添加1.0%时,其抗菌效率达99%,经180 d长期分析,其抗菌活性仍保持95%左右。此外,复合抗菌剂对不同菌株的抗菌性能也均较强。     

Quantitative assessment of the influence of paraoxonase 1 activity and coronary heart disease risk

Human paraoxonase 1 (PON1) is a calcium-dependent high-density lipoprotein associated ester hydrolase that has attracted considerable attention as a candidate factor for coronary heart disease (CHD) based on its function as a key factor in lipoprotein catabolism pathways. This meta-analysis aimed to clarify the inconsistency of published studies and to establish a comprehensive picture of the relationship between PON1 activity and CHD susceptibility. A systematic search was performed from PubMed, Web of Science, EMBASE, and CNKI databases. Ratio of means (RoM) between case and control and 95% confidence intervals (CIs) were calculated using a random-effects model. The source of heterogeneity was explored by subgroup analysis and meta-regression. We identified 47 eligible studies including a total of 9853 CHD cases and 11,408 controls. The pooled analysis showed that CHD patients had a 19% lower PON1 activity than did the controls (RoM=0.81; 95% CI: 0.74-0.89, p<10(-5)). In the subgroup analyses by CHD end points, a similar effect size was observed with coronary stenosis and myocardial infarction subgroups, with corresponding RoM of 0.81 (95% CI: 0.73-0.89, p<10(-4)) and 0.83 (95% CI: 0.74-0.93, p=0.001), respectively. Decreased PON1 activity associated with CHD risk was observed in almost all subgroup analysis according to ethnicity, sample size, study design, mean age of cases, source, and type of control. Decreased PON1 activity may act as a risk factor for the development of CHD. Progressive decrease in serum PON1 activity may exist for an individual with severe disease. However, larger studies using a prospective approach are needed to confirm our results.     

Quantitative genetic tools for insecticide resistance risk assessment: estimating the heritability of resistance

Quantitative genetic studies of resistance can provide estimates of genetic parameters not available with other types of genetic analyses. Three methods are discussed for estimating the amount of additive genetic variation in resistance to individual insecticides and subsequent estimation of the heritability (h2) of resistance. Sibling analysis and offspring-parent regression permit direct estimates of h2 by comparing the resistance phenotypes of individuals of known relatedness. Threshold trait analyses, performed on data from selection experiments, provide estimates of realized heritability. Procedures are outlined for predicting changes in resistance to insecticides based on h2 estimates. Quantitative genetic theory is examined as it relates to resistance and resistance as a quantitative trait; quantitative genetic methods also are unique in providing estimates of genetic correlations between traits. Comments are included on estimates of genetic correlation between resistance and phenotypic traits (e.g., development time) and how they may be used to predict changes in the genetic aspects of phenology that result from insecticide applications (i.e., to predict how the reproductive capacity of future generations will differ from that of the treated generation).     

Quantitative assessment of protein-bound tyrosine nitration in airway secretions from patients with inflammatory airway disease

Because reactive nitrogen species (RNS) have potent inflammatory activity, they may be involved in the inflammatory process in pulmonary diseases. We recently reported increased numbers of 3-nitrotyrosine immunopositive cells, which are evidences of RNS production, in the sputum of patients with chronic obstructive pulmonary disease (COPD) and patients with asthma compared with healthy subjects. In the present study, we attempted to quantify this protein nitration in the airways by means of high-performance liquid chromatography (HPLC) used together with an electrochemical detection system that we developed. Sputum samples were obtained from 15 stable COPD patients, 9 asthmatic patients and 7 healthy subjects by using hypertonic saline inhalation. The values for the molar ratio of protein-bound 3-nitrotyrosine/tyrosine in patients with asthma (4.31 +/- 1.13 x 10(-6), p < 0.05) and patients with COPD (3.04 +/- 0.36 x 10(-6), p < 0.01) were significantly higher than those in healthy subjects (1.37 +/- 0.19 x 10(-6)). The levels of protein-bound 3-nitrotyrosine in the airways were not significantly different in asthmatic patients and COPD patients. A significant negative correlation was found between values for protein-bound 3-nitrotyrosine/tyrosine and % FEV1 values in patients with COPD (r = -0.53, p < 0.05) but not in patients with asthma. These results suggest that our HPLC-electrochemical method is useful for quantifying RNS production in human airways. More importantly, they show that increased RNS production in the airways seems to contribute in a critical way to the pathogenesis of COPD, and that the effects of RNS in airways may differ in asthma and COPD.     

Quantitative assessment of DNA condensation

A fluorescent method is proposed for assessing DNA condensation in aqueous solutions with variety of condensing agents. The technique is based on the effect of concentration-dependent self-quenching of covalently bound fluorophores upon DNA collapse. The method allows a more precise determination of charge equivalency in titration experiments with various polycations. The technique's ability to determine the number of DNA molecules that are condensed together in close proximity is under further investigation.     

Quantitative assessment of anaplerosis from propionate in pig heart in vivo

Normal cardiac metabolism requires continuous replenishment (anaplerosis) of catalytic intermediates of the citric acid cycle. Little is known about the quantitative aspects of propionate as a substrate of in vivo anaplerosis; therefore, we measured the rate of propionate entry into the citric acid cycle in hearts of anesthetized pigs. [U-(13)C(3)]propionate (0.25 mM) was infused in a coronary artery branch for 1 h via an extracorporeal perfusion circuit, and cardiac biopsies were analyzed for the mass isotopomer distribution of citric acid cycle intermediates. Infusion of propionate did not affect myocardial oxygen consumption, heart rate, or contractile function. In the infused territory, propionate infusion did not affect uptake of glucose and lactate but decreased free fatty acid uptake by one-half (P < 0.05). Propionate extraction and uptake were 57.4 +/- 3.3% and 0.078 +/- 0.009 micromol x min(-1) x g(-1). Anaplerosis from propionate, calculated from the mass isotopomer distribution of succinate, accounted for 8.9 +/- 1.3% of the citric acid cycle flux. Propioylcarnitine release accounted for only 0.033 +/- 0.002% of propionate uptake. Methylcitrate did not accumulate. Thus administration of a low concentration of propionate appears to be a convenient and safe way to boost anaplerosis in the heart.     

Quantitative assessment of the influence of PPARG P12A polymorphism on gestational diabetes mellitus risk

The peroxisome proliferator-activated receptor-γ (PPARG) is a member of the nuclear hormone receptor superfamily that has attracted considerable attention as a candidate gene for gestational diabetes mellitus (GDM) based on its function as a key factor involved in the regulation of adipocyte differentiation as well as lipid and glucose metabolism and insulin sensitivity. Many studies have examined the association between P12A polymorphism (rs1801282) in the PPARG gene and risk of GDM, but the results have been inconsistent. To derive a more precise estimation of the relationship, a meta-analysis of 2,858 GDM patients and 6,890 controls from nine published case–control studies was performed. An overall random effects odds ratio of 0.89 (95 % confidence interval [CI]: 0.77–1.04, P = 0.15) was found for 12A allele. In the subgroup analysis by ethnicity, significantly decreased risks were found in East Asians, while no significant associations were detected among Caucasian and Middle Eastern populations. Similar results were also observed using dominant genetic model. This meta-analysis suggested an overall weak association between the P12A polymorphism and GDM risk among East Asians. However, additional very large-scale studies are warranted to provide conclusive evidence on the effects of the PPARG gene on risk of GDM.     

Cytokines in cervicovaginal washing fluid from patients with cervical neoplasia

Human papillomavirus (HPV) infections play an important role in the development of cervical neoplasia. To get to a better understanding of the role of cytokines in the development of these neoplasias, we analysed the presence of various cytokines in cervicovaginal washings of healthy volunteers (n=22), cervical intraepithelial neoplasia (CIN) patients (n=63) and cervical cancer patients (n=33). IL-12p40, IL-10, TGF-beta1, TNF-alpha and IL-1beta levels were significantly higher in patients with cervical cancer than in controls and CIN patients. The levels of IFN-gamma were not different. Our data demonstrate alterations in the local cervical immune environment in cervical cancer patients. This could have important consequences for the further development of immune modulating therapies and vaccination strategies.     

Development of novel lipid-peptide hybrid compounds with antibacterial activity from natural cationic antibacterial peptides

Seven depsipeptides were synthesized by appending seven amino acids (Lys, Leu, Val, Phe, Ser, Gln, and Pro) at the N-terminus of the active fragment [TE-(33-43)], respectively corresponding to the C-terminal beta sheet domain of tenecin 1, an antibacterial protein and their activities were measured against Staphylococcus aureus. Considering the relationship between the activity and the characteristic of amino acid at the N-terminal of the peptide, novel derivatives were designed and synthesized from TE-(33-43) by introduction of fatty acids at the N-terminal. In this process, we synthesized novel lipid-peptide hybrid compounds with a potent antibacterial activity and more improved bioavailabilities. We characterized the important structural parameters of the lipid-peptide hybrid compounds for the antibacterial activities.     

Quantitative assessment of automatic reconstructions of branching systems obtained from laser scanning

Background and Aims

Automatic acquisition of plant architecture is a major challenge for the construction of quantitative models of plant development. Recently, 3-D laser scanners have made it possible to acquire 3-D images representing a sampling of an object''s surface. A number of specific methods have been proposed to reconstruct plausible branching structures from this new type of data, but critical questions remain regarding their suitability and accuracy before they can be fully exploited for use in biological applications.

Methods

In this paper, an evaluation framework to assess the accuracy of tree reconstructions is presented. The use of this framework is illustrated on a selection of laser scans of trees. Scanned data were manipulated by experienced researchers to produce reference tree reconstructions against which comparisons could be made. The evaluation framework is given two tree structures and compares both their elements and their topological organization. Similar elements are identified based on geometric criteria using an optimization algorithm. The organization of these elements is then compared and their similarity quantified. From these analyses, two indices of geometrical and structural similarities are defined, and the automatic reconstructions can thus be compared with the reference structures in order to assess their accuracy.

Key Results

The evaluation framework that was developed was successful at capturing the variation in similarities between two structures as different levels of noise were introduced. The framework was used to compare three different reconstruction methods taken from the literature, and allowed sensitive parameters of each one to be determined. The framework was also generalized for the evaluation of root reconstruction from 2-D images and demonstrated its sensitivity to higher architectural complexity of structure which was not detected with a global evaluation criterion.

Conclusions

The evaluation framework presented quantifies geometric and structural similarities between two structures. It can be applied to the characterization and comparison of automatic reconstructions of plant structures from laser scanner data and 2-D images. As such, it can be used as a reference test for comparing and assessing reconstruction procedures.     

Matrix effects, nonuniform reduction and dispersion in risk assessment for Escherichia coli O157

AIMS: To assess the importance of the variation (or spatial heterogeneity) in the doses of Escherichia coli O157 ingested by individuals when estimating the group risk. METHODS AND RESULTS: Exposure scenarios, which differed in how the same total number of E. coli O157 cells were distributed across a group of persons were simulated and the risks of illness in the population estimated using available dose-response data. Differences in the degree of spatial heterogeneity within routes such as direct contact with sheep faeces, consumption of hamburgers and drinking water could affect the magnitude of the predicted risk by over 1000-fold and contribute to an apparent matrix effect. The effect of barriers, such as cooking, on the spatial heterogeneity of E. coli O157 cells remaining in undercooked hamburgers affects the magnitude of the risk by eightfold. Furthermore, a 100-fold reduction in E. coli O157 concentrations by cooking may reduce the risk of illness from consumption of the burgers by only 12-fold (and not the 100-fold perhaps expected). CONCLUSIONS: The development of microbiological risk assessment (MRA) models for E. coli O157 requires information on the degree of dispersion of faeces within routes such as the environment, food and water, so that the proportion of persons actually exposed to faeces, and hence pathogen, can be quantified. The effect of dispersion is greater for pathogens for which there is greater variation in susceptibility within the population. SIGNIFICANCE AND IMPACT OF THE STUDY: In terms of public health protection, minimizing the dispersion of cattle/sheep faeces reduces the risks to humans by confining exposure to fewer individuals. For MRA, estimating the reduction in risk by a treatment process, e.g. cooking, is complicated by spatial heterogeneity. Regional differences in dispersion (e.g. because of rainfall) may contribute to regional differences in the observed attack rate.