Short Communication: The potential of portable near infrared spectroscopy for assuring quality and authenticity in the food chain, using Iberian hams as an example.

This communication assesses the use of a portable near infrared (NIR) instrument to measure quantitative (fatty acid profile) properties and qualitative ('Premium' and 'Non-premium') categories of individual Iberian pork carcasses at the slaughterhouse. Acorn-fed Iberian pigs have more unsaturated fats than pigs fed conventional compound feed. Recent advances in miniaturisation have led to a number of handheld NIR devices being developed, allowing processing decisions to be made earlier, significantly reducing time and costs. The most common methods used for assessing quality and authenticity of Iberian hams are analysis of the fatty acid composition of subcutaneous fat using gas chromatography and DNA analysis. In this study, NIR calibrations for fatty acids and classification as premium or non-premium ham, based on carcass fat measured in situ, were developed using a portable NIR spectrometer. The accuracy of the quantitative equations was evaluated through the standard error of cross validation or standard error of prediction of 0.84 for palmitic acid (C16:0), 0.94 for stearic acid (C18:0), 1.47 for oleic acid (C18:1) and 0.58 for linoleic acid (C18:2). Qualitative calibrations provided acceptable results, with up to 98% of samples (n = 234) correctly classified with probabilities ⩾0.9. Results indicated a portable NIR instrument has the potential to be used to measure quality and authenticity of Iberian pork carcasses.

Computer image analysis: an additional tool for the identification of processed poultry and mammal protein containing bones.

The aims of this study were (1) to evaluate the potential of image analysis measurements, in combination with the official analytical methods for the detection of constituents of animal origin in feedstuffs, to distinguish between poultry versus mammals; and (2) to identify possible markers that can be used in routine analysis. For this purpose, 14 mammal and seven poultry samples and a total of 1081 bone fragment lacunae were analysed by combining the microscopic methods with computer image analysis. The distribution of 30 different measured size and shape bone lacunae variables were studied both within and between the two zoological classes. In all cases a considerable overlap between classes meant that classification of individual lacunae was problematic, though a clear separation in the means did allow successful classification of samples on the basis of averages. The variables most useful for classification were those related to size, lacuna area for example. The approach shows considerable promise but will need further study using a larger number of samples with a wider range.

The transfer of NIR calibrations for undried grass silage from the laboratory to on-site instruments: comparison of two approaches.

As a first step in a project whose aim is to implement near infrared (NIR) analysis of animal feed on the farm, the present work has examined the possibility of transferring undried grass silage calibrations for dry matter, crude protein, and neutral detergent fiber from a dispersive laboratory NIR instrument (Foss NIRSystem 6500) to a diode array on-site NIR instrument (Zeiss Corona 45 visNIR 1.7). Because the samples are complex and heterogeneous and have high humidity levels it is not easy to establish good calibrations, and it is even more of a challenge to transfer them. By cutting the spectral range to 1100-1650 nm and treating with first or second derivative followed by standard normal variate (SNV) scatter correction, it was possible to obtain very similar spectra from the two instruments. To make the transfer, two approaches were tried. Simply correcting the Corona spectra by subtracting the mean difference spectrum from a transfer set met with only limited success. Making a calibration on the Foss using a calibration set of 503 samples with spectra orthogonalized to the all the difference spectra in the transfer set of 10 samples resulted in a successful transfer for all three calibrations, as judged by performance on two prediction sets of size 22 and 29. Measuring 5 replicate subsamples with the Corona allows it to see a similar surface area to that of 3 replicates in the Foss transport cell, and it is suggested that this is an appropriate level of replication for the Corona.

Improving the discriminatory power of a near-infrared microscopy spectral library with a support vector machine classifier.

A multi-group classifier based on the support vector machine (SVM) has been developed for use with a library of 48,456 spectra measured by near-infrared reflection microscopy (NIRM) on 227 samples representing 26 animal feed ingredients and 4 possible contaminants of animal origin. The performance of the classifier was assessed by a five-fold cross-validation, dividing at the sample level. Although the overall proportion of misclassifications was 27%, almost all of these involved the confusion of pairs of similar ingredients of vegetable origin. Such confusions are unimportant in the context of the intended use of the library, which is the detection of banned ingredients in animal feed. The error rate in discrimination between permitted and banned ingredients was just 0.17%. The performance of the SVM classifier was substantially better than that of the K-nearest-neighbors method employed in previous work with the same library, for which the comparable error rates are 36% overall and 0.39% for permitted versus banned ingredients.

Spectral library validation to identify ingredients of compound feedingstuffs by near infrared reflectance microscopy.

To guarantee feed quality and safety the development and improvement of analytical methods for feed authentication and detection of contaminants is fundamental. Near infrared reflectance microscopy (NIRM) has been investigated as an alternative method to contribute to control systems for feed materials. The major task is the need to build NIRM reference spectral libraries that must represent the variability in feed ingredients. The aim of the present work was to evaluate the performance of a NIRM reference spectral library on animal feed, with external samples of animal feed ingredients and possible contaminants such as processed animal proteins, and in particular to assess its ability to identify ingredients in mixtures. Three external sample sets were used: (A) artificial mixtures, (B) synthetic mixtures and (C) synthetic binary mixtures. The prediction and repeatability results for set A, in which the spectra are from pure ingredients, were very good for both animal and vegetable ingredients and confirm that the spectral library is very good at identifying spectra from pure ingredients. For sets B and C, in which the spectra were measured on mixtures, the prediction results were very disappointing compared with the artificial samples. This means that a strategy that tries to match the spectra taken from a mixture with those of pure ingredients is unlikely to meet with much success. It is possible that an interpolation between pure ingredients for suitably chosen spectral ranges may provide a way to extend this system to mixtures, including mixtures of several ingredients.

Advanced nonlinear approaches for predicting the ingredient composition in compound feedingstuffs by near-infrared reflection spectroscopy.

For quantitative applications, the most common usage of near-infrared reflection spectroscopy (NIRS) technology, calibration involves establishing a mathematical relationship between spectral data and data provided by the reference. This model may be fairly complex, since the near-infrared spectrum is highly variable and contains physical/chemical information for the sample that may be redundant, and multivariate calibration is usually required. When the relationship to be modeled is nonlinear, classical regression methods are inadequate, and more complex strategies and algorithms must be sought in order to model this nonlinearity. The development of NIRS calibrations to predict the ingredient composition, i.e., the inclusion percentage of each ingredient, in compound feeds is a complex task, due to the nature of the parameters to be predicted and to the heterogeneous nature of the matrices/formulas in which each ingredient participates. The present paper evaluates the use of least squares support vector machines (LSSVM) and two local calibration methods, CARNAC and locally biased regression, for developing NIRS models to predict two of the most representative ingredients in compound feed formulations, wheat and sunflower meal, using a large spectral library of 7523 commercial compound feed samples. For both ingredients, the best results were obtained using CARNAC, with standard errors of prediction (SEP) of 1.7% and 0.60% for wheat and sunflower meal, respectively, and even better results when the algorithm was allowed to refuse to predict 10% of the unknowns. Meanwhile, LSSVM performed less well on wheat (SEP 2.6%) but comparably on sunflower meal (SEP 0.60%), giving results very similar to those reported previously for artificial neural networks. Locally biased regression was the least successful of the three methods, with SEPs of 3.3% for wheat and 0.72% for sunflower meal. All the nonlinear methods improved on the standard approach using partial least squares (PLS), which gave SEPs of 5.3% for wheat and 0.81% for sunflower meal.

Measurement error in the explanatory variable of a binary regression: regression calibration and integrated conditional likelihood in studies of residential radon and lung cancer.

In epidemiology, one approach to investigating the dependence of disease risk on an explanatory variable in the presence of several confounding variables is by fitting a binary regression using a conditional likelihood, thus eliminating the nuisance parameters. When the explanatory variable is measured with error, the estimated regression coefficient is biased usually towards zero. Motivated by the need to correct for this bias in analyses that combine data from a number of case-control studies of lung cancer risk associated with exposure to residential radon, two approaches are investigated. Both employ the conditional distribution of the true explanatory variable given the measured one. The method of regression calibration uses the expected value of the true given measured variable as the covariate. The second approach integrates the conditional likelihood numerically by sampling from the distribution of the true given measured explanatory variable. The two approaches give very similar point estimates and confidence intervals not only for the motivating example but also for an artificial data set with known properties. These results and some further simulations that demonstrate correct coverage for the confidence intervals suggest that for studies of residential radon and lung cancer the regression calibration approach will perform very well, so that nothing more sophisticated is needed to correct for measurement error.

Elastic scattering spectroscopy accurately detects high grade dysplasia and cancer in Barrett's oesophagus.

BACKGROUND AND AIMS: Endoscopic surveillance of Barrett's oesophagus currently relies on multiple random biopsies. This approach is time consuming, has a poor diagnostic yield, and significant interobserver variability. Elastic scattering spectroscopy is a real time in vivo optical technique which detects changes in the physical properties of cells. The aim of this study was to assess the potential for elastic scattering to detect high grade dysplasia or cancer within Barrett's oesophagus. METHODS: Elastic scattering spectroscopy measurements collected in vivo were matched with histological specimens taken from identical sites within Barrett's oesophagus. All biopsies were reviewed by three gastrointestinal pathologists and defined as either "low risk" (non-dysplastic or low grade dysplasia) or "high risk" (high grade dysplasia or cancer). Two different statistical approaches (leave one out and block validation) were used to validate the model. RESULTS: A total of 181 matched biopsy sites from 81 patients, where histopathological consensus was reached, were analysed. There was good pathologist agreement in differentiating high grade dysplasia and cancer from other pathology (kappa = 0.72). Elastic scattering spectroscopy detected high risk sites with 92% sensitivity and 60% specificity and differentiated high risk sites from inflammation with a sensitivity and specificity of 79%. If used to target biopsies during endoscopy, the number of low risk biopsies taken would decrease by 60% with minimal loss of accuracy. A negative spectroscopy result would exclude high grade dysplasia or cancer with an accuracy of >99.5%. CONCLUSIONS: These preliminary results show that elastic scattering spectroscopy has the potential to target conventional biopsies in Barrett's surveillance saving significant endoscopist and pathologist time with consequent financial savings. This technique now requires validation in prospective studies.

Impact of process conditions on the centrifugal recovery of a disabled herpes simplex virus.

Despite continuous improvements in culturing and recovery techniques, high-titer stocks of purified disabled herpes simplex virus type-1 (HSV-1 DIS) vector for drug discovery and use in preclinical and clinical trials are currently difficult to achieve. Efforts to improve their centrifugal recovery have been addressed in this paper. The operation of a swing-out centrifuge rotor was assessed, and its operational conditions were defined for the recovery of viable HSV-1 DIS. 80% virus recovery was achieved after 90 min at 26000g. The 20% loss of virus was attributed to damage to the viral envelope by overcompaction of the pellet and impaction with the base of the centrifuge tube. Virus recovery was increased by a further 10% by using a fixed-angle centrifuge rotor operating at 26000g. Plaque assays of recovered HSV-1 DIS gave values on the order of 10(6) pfu/mL, compared to values typically above 10(9) pfu/mL obtained for the replication-competent HSV-1 viron.

A new test for 'sufficient homogeneity'.

Certified reference materials and materials distributed in proficiency testing need to be 'sufficiently homogeneous', that is, the variance in the mean composition of the distributed portions of the material must be negligibly small in relation to the variance of the analytical result produced when the material is in normal use. The requirement for sufficient homogeneity suggests the use of a formal test. Such tests as have been formulated rely on the duplicated analysis of the material from a number of portions, followed by analysis of variance. However, the outcome is not straightforward. If the analytical method used is very precise, then an undue proportion of the materials will be found to be significantly heterogeneous. If it is too imprecise, the test may be unable to detect heterogeneity. Moreover, the Harmonised Protocol Procedure (M. Thompson and R. Wood, Pure Appl. Chem., 1993, 65, 2123) seems to be unduly prone to the rejection of material that is in fact satisfactory. We present a simple new statistical approach that overcomes some of these problems.

Near-infrared spectroscopy for dairy management: measurement of unhomogenized milk composition.

The potential of near-infrared spectroscopy to measure fat, total protein, and lactose contents of unhomogenized milk was studied for use in dairy management, as a new tool for on-line milk analysis in the process of milking. Influence of the spectral region, sample thickness, and spectral data treatment on the accuracy of determination was investigated. Transmittance spectra of 258 milk samples, collected at different stages of the milking process, were obtained with a spectrophotometer (NIRSystems 6500; FOSS-NIRSystems, Silver Spring, MD) in the wavelength range from 400 to 2500 nm with sample thicknesses of 1 mm, 4 mm, and 10 mm. The spectral region and sample thickness were found to be significant factors for milk fat and total protein determination but not the lactose determination. The best accuracy was obtained with the 1100 to 2400 nm region, 1-mm sample thickness, and the first derivative data transformation. For the spectral region from 700 to 1100 nm, close accuracy was obtained for fat with a 10-mm sample and for total protein with a 1-mm sample thickness. The sample thickness did not change significantly the accuracy of lactose determination. Different treatments of spectral data did not improve the calibrations for fat and protein. For the region from 700 to 1100 nm, where inexpensive on-line sensors could be used, the highest positive coefficients for fat were at 930, 968, 990, 1026, 1076, and 1092 nm; for lactose were at 734, 750, 786, 812, 908, 974, 982, and 1064 nm; and for total protein were at 776, 880, 902, 952, and 1034 nm.

Risk of lung cancer associated with residential radon exposure in south-west England: a case-control study.

Studies of underground miners occupationally exposed to radon have consistently demonstrated an increased risk of lung cancer in both smokers and non-smokers. Radon exposure also occurs elsewhere, especially in houses, and estimates based on the findings for miners suggest that residential radon is responsible for about one in 20 lung cancers in the UK, most being caused in combination with smoking. These calculations depend, however, on several assumptions and more direct evidence on the magnitude of the risk is needed. To obtain such evidence, a case-control study was carried out in south-west England in which 982 subjects with lung cancer and 3185 control subjects were interviewed. In addition, radon concentrations were measured at the addresses at which subjects had lived during the 30-year period ending 5 years before the interview. Lung cancer risk was examined in relation to residential radon concentration after taking into account the length of time that subjects had lived at each address and adjusting for age, sex, smoking status, county of residence and social class. The relative risk of lung cancer increased by 0.08 (95% CI -0.03, 0.20) per 100 Bq m(-3) increase in the observed time-weighted residential radon concentration. When the analysis was restricted to the 484 subjects with lung cancer and the 1637 control subjects with radon measurements available for the entire 30-year period of interest, the corresponding increase was somewhat higher at 0.14 per 100 Bq m(-3) (95% CI 0.01, 0.29), although the difference between this group and the remaining subjects was not statistically significant. When the analysis was repeated taking into account uncertainties in the assessment of radon exposure, the estimated increases in relative risk per 100 Bq m(-3) were larger, at 0.12 (95% CI -0.05, 0.33) when all subjects were included and 0.24 (95% CI -0.01, 0.56) when limited to subjects with radon measurements available for all 30 years. These results are consistent with those from studies of residential radon carried out in other countries in which data on individual subjects have been collected. The combined evidence suggests that the risk of lung cancer associated with residential radon exposure is about the size that has been postulated on the basis of the studies of miners exposed to radon.

Leukemia mortality after X-ray treatment for ankylosing spondylitis.

Leukemia mortality has been studied in 14,767 adult ankylosing spondylitis patients diagnosed between 1935 and 1957 in the United Kingdom, of whom 13,914 patients received X-ray treatment. By 1 January 1992, there were 60 leukemia deaths among the irradiated patients, almost treble that expected from national rates. Leukemia mortality was not increased among unirradiated patients. Among those irradiated, the ratio of observed to expected deaths for leukemia other than chronic lymphocytic leukemia was greatest in the period 1-5 years after the first treatment (ratio = 11.01, 95% confidence interval 5.26-20.98) and decreased to 1.87 (95% confidence interval 0.94-3.36) in the 25+ year period. There was no significant variation in this ratio with sex or age at first treatment. The ratio for chronic lymphocytic leukemia was slightly but not significantly raised (ratio = 1.44, 95% confidence interval 0.62-2.79). Most irradiated patients received all their exposure within a year. Based on a 1 in 15 random sample, the mean total marrow dose was 4.38 Gy. Doses were nonuniform, with heaviest doses to the lower spine. The risk for nonchronic lymphocytic leukemia was adequately described by a linear-exponential model that allowed for cell sterilization in heavily exposed parts of the marrow and time since exposure. Ten years after first exposure, the linear component of excess relative risk was 12.37 per Gy (95% confidence interval 2.25-52.07), and it was estimated that cell sterilization reduced the excess relative risk by 47% at 1 Gy (95% confidence interval 17%-79%). The average predicted relative risk in the period 1-25 years after exposure to a uniform dose of 1 Gy was 7.00.

Comparison of cervical magnetic stimulation and bilateral percutaneous electrical stimulation of the phrenic nerves in normal subjects.

Cervical magnetic stimulation is a new technique for stimulating the phrenic nerves, and may offer an alternative to percutaneous electrical stimulation for assessing diaphragmatic strength in normal subjects and patients in whom electrical stimulation is technically difficult or poorly tolerated. We compared cervical magnetic stimulation with conventional supramaximal bilateral percutaneous electrical stimulation in nine normal subjects. We measured oesophageal pressure (Poes), gastric pressure (Pgas) and transdiaphragmatic pressure (Pdi). The maximal relaxation rate (MRR) was also measured. The mean magnetic twitch Pdi was 36.5 cmH2O (range 27-48 cmH2O), significantly larger than electrical twitch Pdi, mean 29.7 cmH2O (range 22-40 cmH2O). The difference in twitch Pdi was explained entirely by twitch Poes, and it is possible that the magnetic technique stimulates some of the nerves to the upper chest wall muscles as well as the phrenic nerves. We compared bilateral, rectified, integrated, diaphragm surface electromyographic (EMG) responses in three subjects and found results within 10% in each subject, indicating similar diaphragmatic activation. The within occasion coefficient of variation, i.e. same subject/same session, was 6.7% both for magnetic and electrical twitch Pdi. The between occasion coefficient of variation, i.e. same subject/different days, was 6.6% for magnetic stimulation and 8.8% for electrical. There was no difference between relaxation rates measured with either technique. We conclude that magnetic stimulation is a reproducible and acceptable technique for stimulating the phrenic nerves, and that it provides a potentially useful alternative to conventional electrical stimulation as a nonvolitional test of diaphragm strength.

Patterns of lung cancer mortality among uranium miners in West Bohemia with varying rates of exposure to radon and its progeny.

Lung cancer mortality in a cohort of 4320 miners first employed during 1948-1959 at the Jáchymov and Horní Slavkov uranium mines in West Bohemia and followed until 1 January 1991 has been studied to gain a greater understanding of the consequences of exposure to radon and its progeny. Among men whose exposure rates never exceeded 10 working levels, excess relative risks per unit exposure were greater in younger men, and exposures received in the periods 15-24, 25-34 and 35+ years previously were found to have 47, 24 and 0% of the effect of exposures 5-14 years previously. Within this low-exposure-rate group excess relative risk increased linearly with time-weighted cumulative exposure and did not depend on exposure rate or duration of exposure. For men who spent less than 20% of their employment at the Jáchymov mine the excess relative risk per working level month was 1.36% (95% confidence interval 0.52-3.54) in the baseline category (age group 55+ and exposure received 5-14 years previously). For men who spent more than 20% of their employment at Jáchymov, the corresponding excess relative risk per working level month was higher by a factor of 1.80 (95% confidence interval 1.27-2.97). The difference may be due to the fact that men who spent more than 20% of their employment at Jáchymov were exposed to the much higher levels of arsenic in the dust at the Jáchymov mine than at other mines. When men with exposure rates above 10 working levels were included in the analysis, patterns of risk were complex and depended on both exposure rate and duration of exposure in addition to the factors mentioned above. If these findings are confirmed elsewhere, calculation of risk estimates for extrapolation to modern occupational or environmental exposures should be based on miners with exposure rates below about 10 working levels. Further investigation is desirable of the influence of dusts containing arsenic on lung cancer risk in miners exposed to radon.

A low-cost system for computerized rodent weight logging with statistical analysis.

A system comprising a suite of 4 computer programs has been developed for on-line rodent weight data collection and statistical analysis using an IBM personal computer. Data can be collected from up to 3 separate trials simultaneously, and can be stored for later statistical analysis. Mettler balances were used for the animal weighing. A Mettler current loop adapter and a multiplexer were used to interface the balances with the computer.

Cereal dietary fiber consumption and diverticular disease: a lifespan study in rats.

The relationship between consumption of dietary fiber (DF) from white bread, wholemeal bread, or bran and the development of diverticular disease of the colon has been investigated in a lifespan study using 1800 Wistar rats in nine diet groups. Use of the rat as a model for the human condition was validated by demonstration of significant relationships between fiber intake and fecal output and transit time, and the observation of true acquired diverticula, both single and multiple. Significant inverse relationships (mostly with p less than 0.001) were observed between the incidence of diverticula (and prediverticula) and the concentrations of fiber in the diets, measured by the neutral detergent fiber and Southgate methods. The study offers strong support to the Painter-Burkitt view of human diverticular disease as being due to fiber deficiency, if the extrapolation from rat to man is valid. On the same assumption, the amount of additional fiber required to be consumed in order to achieve a substantial reduction in incidence of the disease is very large. Effects of fiber on body weight, food intake, mineral levels, blood composition and properties, mortality, organ weights, and incidence of tumors and lesions are reported. Significantly fewer mammary tumors were found in rats fed the very high fiber stock diet than in those fed the purified diets.

The Chatham Blood Pressure Study. An application of Bayesian growth curve models to a longitudinal study of blood pressure in children.

Recent developments in statistics have produced powerful methods that facilitate the analysis of longitudinal studies. These methods are illustrated by an analysis of a longitudinal study of blood pressure in children. The results of the study show a clear tendency for blood pressure to increase with age, and Asian children tend to have lower blood pressures than their Caucasian counterparts of the same age. There is evidence to support the hypothesis that blood pressures track.