The image analysis of colour changes of different human tissues in the relation to the age. Part 2: practical applicability.

The method of image analysis of intervertebral disc, Achilles tendon and rib cartilage was applied for assessment of colour changes of these tissues in the relation to the human age. It was proved that colour of tested tissues changes with age which is most obvious on rib cartilage and intervertebral disc, while Achilles tendon does not display important changes. The parameters MeanBlue, MeanSaturation and MeanBrightness are the best for age estimation based on colour analysis.

The image analysis of colour changes of different human tissues in the relation to the age. Part 1. Methodological approach.

The human age for medico-legal purposes is usually estimated from hard tissues like bones and teeth. Only little attention was paid to soft tissues most probably due to the lack of detectable age changes. This study deals with colour changes of human tissue from intervertebral discs, Achilles tendon and rib cartilage in the relation to the age. The image analysis of colour of investigated tissue samples was performed. The values of intensities of channels RGB (MeanRed, MeanGreen, and MeanBlue) and parameters from the IHS system (MeanSaturation, HueTypical, HueVariation, BrightVariation and MeanBrightness) were evaluated. The results confirm that colour changes of some tissues can be used for age estimation.

Contents of pentosidine in the tissue of the intervertebral disc as an indicator of the human age.

The study deals with the post-translational modifications of proteins - glycation of the tissue of the intervertebral disc and determination of one of advanced glycation end's products - pentosidine in the relation to the age. Pentosidine was detected in the hydrolysate of the intervertebral discs from persons between the ages of 16 and 95 years. 142 samples were analysed by high performance liquid chromatography, and the detected amounts of pentosidine were processed statistically. The coefficient of correlation of dependence of the amount of pentosidine on the age amounts to r = 0.92. The results of the work testify to the fact that it is possible to use the detection of pentosidine in the tissue of the intervertebral disc for the estimation of the age. Nevertheless subsequent experiments should be done under different conditions post-mortem decomposition.

[Age-related post-translational modification of proteins in human tissue and its use determination of age in forensic medicine. Review]. / Závislost nekterých posttranslacních modifikací proteinu lidských tkání na veku a jejich pouzití pro urcování veku v soudním lékarství. Prehled poznatku.

This article deals with some of posttranslational modification of proteins which are, or could be useful to work out objective and standard methods for age estimation in forensic medicine. From many posttranslational modifications other than racemization, the evaluation of pentosidine, one of the products of nonenzymatic glycosylation, seems to be promising. The enlargement of menu of methods for age estimation is important mainly for forensic sciences when determination of the age of an unknown dead body is necessary. Morphological methods are quite often subjective and charged with errors.

The use of the D-, L- aspartic ratio in decalcified collagen from human dentin as an estimator of human age.

Among the methods dealing with the age estimation, the evaluation of the ratio of the D-, L- form of the aspartic acid in tissues with a low metabolic turnover is considered to be the most precise. We introduced demineralization of the dentin with 0.5 M EDTA adjusted to pH = 7.4. The advantage of such a procedure is that after demineralization we obtained pure insoluble protein (collagen) and soluble noncollagenous proteins in one step. In this study we analyzed insoluble collagen. The amino acids obtained after the hydrolysis were derivatized into TFA isopropyl esters and analyzed by gas chromatography on Chirasil L-Val capillary column. We analyzed human dentin from the lower canines. The correlation coefficient was 0.93 for our set of 71 persons. The result concurred with those of other scientists.

[Determination of the D-and L-forms of aspartic acid in intervertebral disk tissue as an indicator of age]. / Stanovení pomeru D- a L-formy kyseliny asparagové ve tkáni intervertebrálního disku jako ukazatel veku.

Assessment of age according to racemization coefficient of aspartic acid in dentin is considered a reliable method. Proteins which are isolated from dentin are non-collagenous proteins or collagen. As collagen is one of the most frequent proteins in the human body, the question arises whether the D-form of aspartic acid can be assessed also in other tissues and the racemization coefficient used for assessment of age. In the submitted work the author assessed to use the racemization coefficient in the tissue of the intervertebral disk. A lyophilized portion of the annulus fibrosus was hydrolyzed and the amino acids of the hydrolysate were estimated after derivation by gas chromatography on a chiral capillary column. The values of the racemization coefficient indicate limited possibilities to assess age from thus prepared tissue.

[Determination of D- and L- forms of aspartic acid in Achilles tendon tissue as an indicator of age]. / Stanovení pomeru D- a L-formy kyseliny asparagové ve tkáni Achillovy slachy jako ukazatel veku.

The presence of the D-form of aspartic acid was detected in collagen of dentin and the intervertebral disk before and assessment of its racemization coefficient was used for assessment of the age of subjects. Another tissue formed almost solely by collagen type I are tendons. Tendon tissue was not investigated so far with regard to the racemization coefficient and therefore the authors assessed the D-form of aspartic acid in the Achilles tendon. They used lyophilized tendon excisions which were hydrolyzed. Amino acids in the hydrolysate were derived to N-TFA isopropyl esters which were assessed by gas chromatography on a chiral capillary column. In all samples the D-form of aspartic acid was detected. The values of the racemization coefficient suggest the possible use of thus processed tissue with certain limitations.

Effect of amino acids on the composition and properties of extruded mixtures of wheat flour and glucose.

Wheat flour was extruded at 100-120 degrees C with 5% D-glucose or mixtures of 5% D-glucose and 0.5% or 2.0% of L-alanine, L-leucine, L-lysine, L-threonine or L-cysteine. The extent of browning was only moderate, and yellow and red pigments were produced. The odour intensity increased with the addition of either glucose or a mixture of glucose and amino acids; the odour was not significantly intensive according to sensory acceptancy analysis. The odour profile was influenced by the structure of amino acid present in the extruded material. The addition of D-glucose increased the production of furan derivatives, and in lesser degree, of pyrazines in the extruded product. The pyrazine content increased with the addition of amino acids, except cysteine which enhanced the production of sulphur compounds instead of pyrazines. The composition of the pyrazine fraction varied depending on the amino acid added. Pretreatment of D-glucose with an amino acid in aqueous solution affected the composition of volatiles, but it did not significantly enhance the pyrazine production.

Determination of the size of a foreign body in the eye using image analysis of its roentgenogram.

The case of a computer-enhanced image analysis of X-ray pictures of a pellet in the orbit of a man who sustained a shotgun injury to his right eyeball is presented. The man was shot accidentally during a pheasant hunt. Two hunters were shooting simultaneously but they were using three different sizes of pellet--3.0, 3.5 and 4.0 mm. One of the pellets hit a third man in the eye. He sustained serious injury with a resultant loss of sight in this eye despite immediate medical treatment. Unfortunately the pellet could not be removed from the orbit, but it was necessary to establish who was responsible for the injury. The problem lay in determining the specification and size of the pellet at that moment. The problem was solved using computer-enhanced video image analysis of the X-ray pictures of the pellets. The image processor LUCIA G (http(/)/www.lim.cz) was used.

[Age determination by detection of ratios of the D and L forms of aspartic acid]. / Urcení veku stanovením pomeru D,L formy kyseliny asparagové.

Paper concerns evaluation of results of age estimation based on the increase of D-aspartic acid during aging. Samples of dentin from lower canine teeth were used as experimental material. In addition, analytical pitfalls and new possibilities of d-amino acids detection were discussed.

Prediction of odour and flavour acceptances of white wines on the basis of their colour.

The colour of Moravian white wines was evaluated instrumentally, using the trichromatic method, and by sensory analysis. The odour and flavour acceptances could be predicted very well on the basis of sensory colour evaluation, and less efficiently by instrumental method. Consumers preferred wines with prevailing yellow hue, which was associated with sweet, fruity and floral flavour notes. Green hue was considered a negative factor, without any rational relation to negatively perceived flavour notes. Logarithmic relations fit moderately better experimental results than linear relations.

[Age determination by measurement of the ratio of D- and L- forms of aspartic acid. II. Chiral separation of amino acids from human dentin]. / Moznosti urcení veku stanovením pomeru D, L-formy kyseliny asparagové II. Chirální separace aminokyselin z lidského dentinu.

Estimation of relation of D,L-forms aspartic acid has been recently used for an evaluation of age. It represents a relatively new method based on analysis of hydrolyzed dentine. Its amino acids are derived and analyzed by gas chromatography or liquid chromatography on a chiral column.

[Present possibilities of age determination in forensic medicine with emphasis on the importance of measurement of D- and L- forms of aspartic acid. I. An overview]. / Soucasné moznosti urcení veku v soudne-lékarské praxi se zamerením na význam stanovení D, L-forem kyseliny asparagové I. Prehled poznatku.

Evaluation of age of unknown deceased persons belongs to the most important ways to identification. For the time being, morphological methods are used, namely evaluation of age according to Gustafson's method from tooth grindings or by macroscopical estimation of abrasion, transparency of root dentine, alveolar atrophy and number of missing teeth. Evaluation of the data can be influenced by an individual failure and experience showed an age related decrease of precision. Recently, some papers occurred estimating a relation of D, L-forms of aspartic acid which depends on the age with a significant precision.

Determination of aryloxyphenoxypropanoic acid derivatives in crops treated with herbicidal sprays.

A reproducible and sensitive gas chromatographic method has been developed for the trace analysis of fluazifop-butyl, haloxyfop-ethoxyethyl and quizalofop-ethyl as well as their main metabolites (corresponding acids analysed after conversion to methyl esters) in some fruits and vegetables treated with herbicidal sprays. Comparable detection limits (0.01 mg/kg) were achieved when mass fragmentography and/or the electron capture detector were employed. However, in the latter case, bromination of fluazifop esters had to be carried out prior to GC analysis. By means of the nitrogen-phosphorus detector, determination of residues at a level of 0.05 mg/kg was possible. The residue values found in strawberries, cabbages, cauliflower and carrots were used to discuss the degree of metabolic hydrolysis of parent esters, i.e. active ingredients of herbicides.

Gas chromatographic determination of chlorinated phenols in the form of various derivatives.

Derivatization of chlorophenols prior to gas chromatographic analysis resulted in a significant improvement of their chromatographic behaviour. Chlorophenols in the form of anisoles, acetates, and pentafluorobenzyl ethers were separated either on a HP-1 wide-bore capillary column or on a capillary column coated with a film of SE-54. Due to an enhancement of the poor electron-capture detection response of mono- and dichlorophenols, pentafluorobenzylation made it possible to determine trace amounts in spiked water. Moreover, simultaneous analysis of phenoxyalkanoic acids was accomplished under the same conditions.

Gas chromatographic-mass spectrometric investigation of phenoxypropanoic acid derivatives possessing herbicidal activity.

Fluazifop-butyl, haloxyfop-ethoxyethyl and quizalofop-ethyl, active ingredients of new selective herbicidal preparations, as well as their metabolites converted in to corresponding methyl esters were separated on several gas chromatographic columns. Bromination of fluazifop esters increased significantly the response of the electron-capture detector. Electron impact mass spectra of all the compounds investigated were recorded, and characteristic ions suitable for selected ion monitoring are given.

Residues of clopyralide (3,6-dichloropicolinic acid) in strawberries and their changes.

Residues of clopyralide in strawberries treated with herbicide preparation Lontrel 300 were determined using gas chromatographic method. The decrease of clopyralide content during the processing of contaminated fruits was observed. The rate of decomposition of this compound in model solutions (held at 75 degrees C, pH 3.6) followed first-order kinetics. 2,5-dicloropyridine was proved to originate under these conditions from parent compound.