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Manipulation of protein fingerprints during on-column fluorescent labeling: protein fingerprinting of six Staphylococcus species by capillary electrophoresis.

Bacterial proteomes were analyzed by use of electrophoretically mediated microanalysis (EMMA) and field-enhanced stacking. A water-soluble protein fraction was injected onto a capillary. Next, a fluorogenic reagent was injected and allowed to react with the protein mixture, producing fluorescent products that were separated by submicellar capillary electrophoresis and detected by laser-induced fluorescence. By use of a low-ionic strength sample buffer and a brief electrophoretic step, slow moving anionic proteins were stacked at the reagent-sample interface and were preferentially labeled. By reversing the order of sample injection and labeling reagent, fast moving cationic proteins were preferentially labeled. By adjustment of the sample buffer pH, proteins with different isoelectric points were selectively labeled. Electrophoresis fingerprints were generated for the water-soluble protein fraction from six Staphylococcus species. The protein patterns produced were species-specific and were used to construct a phylogenetic tree.

Bacterial Proteins↗

Pharmaceutical fingerprinting in phase space. 1. Construction of phase fingerprints.

The present study proposes a general method for constructing pharmaceutical fingerprints in the analysis of HPLC trace organic impurity patterns. The approach considers signals in phase space and accounts for two different types of noise: additive and perturbative. The first type, additive noise, contributes to distortion of the absolute values of signal peaks. The second type, perturbative noise, contributes to variations of the retention times of signal peaks and distorts the time scale of the trace organic impurity patterns. The ability of the proposed approach to consider both types of noise significantly distinguishes it from existing methods of data analysis that are usually designed to treat only the additive noise. Analysis of the HPLC signals in phase space eliminates the problem of perturbation noise and enables detection and comparison of similar signal segments recorded at different retention times. The current study analyzes the chromatographic trace organic impurity patterns collected from six different manufacturers of L-tryptophan using three HPLC columns. For five manufacturers the variability of data recorded with the same column are in perfect agreement with the proposed model. A significant variance of parameters is detected for one manufacturer, thus indicating a possible change in its product consistency. The analysis in phase space is also used to explain the previously detected variability of HPLC signals across columns. The accompanying paper reports an application of the proposed approach for the pattern recognition of HPLC data.

Algorithms↗

[ABO blood grouping of fingerprint by means of immunohistochemical procedure].

For the purpose of ABO-blood typing on fingerprints, the detection of blood group substances in fingerprints attached on nitrocellulose filter or paper was performed immunohistochemically using avidin-biotin-peroxidase complex (ABC) method. At first, it was fundamentally tested whether ABO-blood typing could be specifically performed for the fingerprints of known ABO blood group, being made experimentally on nitrocellulose filter or paper, and the effect of fixation and paper quality for the detection was also examined. And, ABO-typing was carried out using transferred fingerprints from a slide glass to a nitrocellulose filter. Moreover, using a fingerprint of unknown ABO-type, serially repetitive blood typing (three times) was compared to individually performed grouping (three tests) after dividing a single fingerprint into three parts. In addition, the method of serial ABO-blood typing after the morphological detection of fingerprints by ninhydrine was also considered. As results, according to primary antibodies applied, ABH activities were specifically detected in the fingerprints on nitrocellulose filter and paper. For the fixation procedure of very minute blood group substances to paper, heat fixation was the most effective and methanol fixation was also available. The intensity of immunostaining of fingerprints decreased according to the deterioration of paper quality used. And, the transferred fingerprints on nitrocellulose filter were also specifically typed. Serially repetitive blood typing (anti-B-->anti-A-->anti-H) was possible to fingerprints on nitrocellulose filter, but it gave poor results to those on paper. To overcome this difficulty, after being detected morphologically by ninhydrine, iodine or aluminium powder and decolored thereafter, a fingerprint on paper was divided into three parts and specific blood typing was possible. As for the double blind test to fingerprints on paper, 22 out of 35 fingerprints were specifically typed and the rate of correct typing was 62.9%. From these results, it can be considered that this method is available for ABO-typing of fingerprints of secretor type in forensic practice.

ABO Blood-Group System↗

Identifying the origin of single corneal cells by DNA fingerprinting: part I--implications for corneal limbal allografting.

PURPOSE: To demonstrate that the combination of impression cytology and single cell DNA fingerprinting represents a powerful tool that is suitable for detecting transplanted cells after corneal limbal allografting. METHODS: Fifty single cells were obtained by corneal impression cytology from 12 patients undergoing cataract surgery. Individual cells were isolated from samples by micromanipulation. Polymerase chain reaction and short tandem repeat profiling was used to obtain forensic standard "DNA fingerprints" from single cells. Blood samples taken at the time of impression cytology provided control "fingerprints." RESULTS: Informative DNA fingerprints were obtained from all corneal samples and 66% (33 of 50 cells) of isolated single cells. Of all fingerprints obtained, most (91%, 30 of 33 fingerprints) corneal fingerprints matched corresponding blood sample fingerprints. At least one corneal fingerprint matched the corresponding blood sample fingerprint in 83% (10 of 12 patients) of the patients in the study. CONCLUSIONS: This extremely specific single cell DNA fingerprinting system permits accurate identification of individual corneal epithelial cells, allowing very reliable determination of their origin, which will enable host and donor cells to be distinguished from each other after keratolimbal allografting procedures, even if the host and donor are the same sex or siblings. These DNA fingerprinting methods allow assessment of quality and quantity of donor cell survival, as well as survival time. The extreme sensitivity and accuracy of the technique means that should contamination occur, it would be identified, thus ensuring meaningful results.

Cell Transplantation↗

Pseudo-outer product based fuzzy neural network fingerprint verification system.

Fingerprint identification has been used in law enforcement applications over the last century, and has become the de facto international standard for positive identification. With the emergence of automated fingerprint identification technologies, it has assisted in making the once labour-intensive process of classifying, searching and matching a thing of the past. As a biometrics proof of identification, not many have ventured into the world of fingerprint identification using fuzzy neural networks. In this paper, a database of fingerprint images is constructed and a fuzzy neural network called the pseudo outer product fuzzy neural network (POPFNN) [Zhou, R.W. & Quek, C. (1996). A pseudo outer-product based fuzzy neural network. Neural Networks, 9(9), 1569-1581] is trained to detect similarity between two fingerprints and decide whether they belong to the same person. The fundamental idea is that, given a person's fingerprints taken under different conditions, the POPFNN based fingerprint verification system should be sufficiently robust to distinguish the difference. The people providing the fingerprint samples are subjected to different 'adverse' conditions; from wetness to chemical treatments. Fingerprint images are taken after conditions such as: after a shower, holding pineapples (mild acid from fruit), after washing one's hands, etc. The characteristics of POPFNN, such as the learning, generalisation, and high computational abilities, make fingerprint verification particularly powerful when verifying authentic fingerprints subjected to external conditions and recognising spurious ones. In order to demonstrate the efficacy of POPFNN and its application in the fingerprint verification system (FVS), several types of experiments have been designed and implemented in this work. The experimental results and analysis are presented at the end of the paper for discussion.

Algorithms↗

[Comparison of fingerprints using a personal computer and graphic software sold on market].

We compared the fingerprint images using a personal computer and a graphic software sold on market. The fingerprints of thumb of left hand recorded by pressing and rolling on paper were input in computer using image scanner. These two fingerprint images were overlapped by moving and rotation on the computer screen, and were fitted well. This result showed that the fingerprints recorded by any way from one person were fitted. Latent and accused's fingerprints were also input in computer using image scanner. These images were analyzed by the same procedure. These two fingerprints were part by unfitting. It was decided these two fingerprints were not derived from one person. We also considered the possibility that the latent fingerprint was the reversed one. Then, the image was reversed of its color on the screen of computer (black to white and white to black) and compared by the same procedure. In this experiment, two fingerprint images were not fitted in a part. This showed that the reversed fingerprint was also able to be analyzed by this procedure. For the comparison of fingerprints, this report showed the ability of the personal computer and software which can be obtained easily from market. Following the improvement of the hardware and software used in this examination in near future, the computerized identification of fingerprints like this study will become more accurate and convenient.

Computer Graphics↗

Near homogeneity of PR2-bias fingerprints in the human genome and their implications in phylogenetic analyses.

Genes of a multicellular organism are heterogeneous in the G+C content, which is particularly true in the third codon position. The extent of deviation from intra-strand equality rule of A = T and G = C (Parity Rule 2, or PR2) is specific for individual amino acids and has been expressed as the PR2-bias fingerprint. Previous results suggested that the PR2-bias fingerprints tend to be similar among the genes of an organism, and the fingerprint of the organism is specific for different taxa, reflecting phylogenetic relationships of organisms. In this study, using coding sequences of a large number of human genes, we examined the intragenomic heterogeneity of their PR2-bias fingerprints in relation to the G+C content of the third codon position (P3). Result shows that the PR2-bias fingerprint is similar in the wide range of the G+C content at the third codon position (0.30-0.80). This range covers approximately 89% of the genes, and further analysis of the high G+C range (0.80-1.00), where genes with normal PR2-bias fingerprints and those with anomalous fingerprints are mixed, shows that the total of 95% of genes have the similar finger prints. The result indicates that the PR2-bias fingerprint is a unique property of an organism and represents the overall characteristics of the genome. Combined with the previous results that the evolutionary change of the PR2-bias fingerprint is a slow process, PR2-bias fingerprints may be used for the phylogenetic analyses to supplement and augment the conventional methods that use the differences of the sequences of orthologous proteins and nucleic acids. Potential advantages and disadvantages of the PR2-bias fingerprint analysis are discussed.

Base Composition↗

Fingerprinting of mixed bacterial strains and BIOLOG gram-negative (GN) substrate communities by enterobacterial repetitive intergenic consensus sequence-PCR (ERIC-PCR).

PCR-based genomic fingerprinting by use of enterobacterial repetitive intergenic consensus primers (ERIC-PCR) was evaluated for its use in fingerprinting DNA of mixed Gram-negative bacterial strains and BIOLOG Gram-negative (GN) microplate substrate communities. ERIC-PCR fingerprints of six different pure bacterial strains and a combined mixture of the strains were compared with fingerprints obtained by two more established methods: amplified ribosomal DNA restriction analysis (ARDRA) and random amplified polymorphic DNA analysis (RAPD-PCR). The ERIC-PCR fingerprint of the mixed strains was highly reproducible and was more species-specific and representative of the individual strain fingerprints than the ARDRA and RAPD-PCR fingerprints, respectively. ERIC-PCR fingerprinting of model and rhizosphere BIOLOG GN substrate communities also provided clearly distinguishable fingerprints. Results of this study suggest that ERIC-PCR represents a rapid and highly discriminating method for fingerprinting DNA of mixed Gram-negative bacterial strains and BIOLOG GN substrate communities.

DNA Fingerprinting↗

Improved repetitive-element PCR fingerprinting of Salmonella enterica with the use of extremely elevated annealing temperatures.

Modified thermal cycling conditions were explored in an effort to improve the reproducibility and resolving power of repetitive-element PCR (rep-PCR) fingerprinting. Assay performance was rigorously evaluated under standard and modified cycling conditions, using as a test set 12 strains putatively representing 12 serovars of Salmonella enterica. For all three fingerprint types (ERIC2, BOXA1R, and composite fingerprints), the use of extremely elevated annealing temperatures plus an initial "touchdown" cycling routine yielded significant improvements in day-to-day reproducibility and discriminating power despite the somewhat sparser appearance of the fingerprints. Modified cycling conditions markedly reduced the variability of fingerprints between cyclers, allowing fingerprints from different cyclers to be analyzed together without the degradation of assay performance that occurred with between-cycler analyses under standard cycling conditions. With modified cycling, composite fingerprints exhibited the lowest reproducibility but the highest net discriminating power of the three fingerprint types. rep-PCR fingerprints led to the discovery of a serotyping error involving one of the 12 test strains. These data demonstrate that modified cycling regimens that incorporate elevated annealing temperatures (with or without an initial touchdown routine) may markedly improve the performance of rep-PCR fingerprinting as a bacterial typing tool.

DNA Fingerprinting↗

Comparison of DNA fingerprints and somatic serotypes of serogroup B and E Pasteurella multocida isolates.

The DNA fingerprint profiles and somatic serotypes of 71 Pasteurella multocida capsule serogroup B isolates, 13 capsule serogroup E isolates, and 16 somatic reference serotype strains were compared. Each of the 16 reference somatic serotypes had a unique DNA fingerprint profile with the HhaI restriction endonuclease. Fifty-four serogroup B isolates (isolated from classical cases of hemorrhagic septicemia) reacted with somatic serotype 2 or 5 antiserum and had DNA fingerprint profiles which resembled that of the serotype 2 reference strain. Seven DNA fingerprint profiles were found among 16 serogroup B strains representing other somatic serotypes. The DNA fingerprints of these isolates were different from the fingerprints of the 16 somatic reference serotype strains. All 13 serogroup E isolates had identical somatic serotypes and identical DNA fingerprint profiles when the HhaI endonuclease was used. The HhaI fingerprint profile of the serogroup E isolates did not match any fingerprint profile of the reference somatic serotype strains. Following DNA profiling with the HhaI endonuclease, the 13 serogroup E isolates were differentiated sequentially with HpaII restriction endonuclease. A descriptive identification epithet for P. multocida isolates was constructed. The descriptive epithet consists of serologic identification and sequential DNA profiles with restriction endonucleases HhaI and HpaII, respectively. DNA fingerprinting of P. multocida is a precise characterization method. In conjunction with serologic typing, it can further classify P. multocida isolates for epidemiologic studies.

Animals↗

A format for databasing and comparison of AFLP fingerprint profiles.

BACKGROUND: Amplified fragment length polymorphism (AFLP) is a PCR-based technique that involves restriction of genomic DNA followed by ligation of adaptors to the fragments generated and selective PCR amplification of a subset of these fragments. The amplified fragments are separated on a sequencing gel and visualized by autoradiography or fluorescent sequencing equipment. AFLP allows high-resolution genotyping but the lack of a format for databasing and comparison of AFLP fingerprint profiles limits its wider applications in profiling large numbers of biological samples. RESULTS: A scheme is described to represent a DNA fingerprint profile with a nucleotide sequence-like format in which the information line contains the minimal necessary details to interpret an AFLP DNA fingerprint profile. They include technique used, information on restriction enzymes, primer combination, biological source for DNA materials, fragment sizing and annotation. The bodylines contain information on size and relative intensity of DNA fragments by a string of defined alphabets or symbols. Algorithms for normalizing raw data, binning of fragments and comparing AFLP DNA fingerprint profiles are described. Firstly, the peak heights are normalized against their average and then represented by five symbols according to their relative intensities. Secondly, a binning algorithm based loosely on common springs and rubber bands is applied, which positions sequence fragments into their best possible integer approximations. A BLAST-like reward-penalty concept is used to compare AFLP fingerprint profiles by matching peaks using two metrics: score and percentage of similarity. A software package was developed based on our scheme and proposed algorithms. Example of use this software is given in evaluating novelty of a new tropical orchid cultivar by comparing its AFLP fingerprint profile against those of related commercial cultivars in a database. CONCLUSIONS: AFLP DNA fingerprint profiles can be databased and compared effectively with software developed based on our scheme and algorithms. It will facilitate wider use of this DNA fingerprinting technique in areas such as forensic study, intellectual property protection for biological materials and biodiversity management. Moreover, the same concepts can be applied to databasing and comparing DNA fingerprint profiles obtained with other DNA fingerprint techniques.

Algorithms↗

Forensic analysis of nonlinear collusion attacks for multimedia fingerprinting.

Digital fingerprinting is a technology for tracing the distribution of multimedia content and protecting them from unauthorized redistribution. Unique identification information is embedded into each distributed copy of multimedia signal and serves as a digital fingerprint. Collusion attack is a cost-effective attack against digital fingerprinting, where colluders combine several copies with the same content but different fingerprints to remove or attenuate the original fingerprints. In this paper, we investigate the average collusion attack and several basic nonlinear collusions on independent Gaussian fingerprints, and study their effectiveness and the impact on the perceptual quality. With unbounded Gaussian fingerprints, perceivable distortion may exist in the fingerprinted copies as well as the copies after the collusion attacks. In order to remove this perceptual distortion, we introduce bounded Gaussian-like fingerprints and study their performance under collusion attacks. We also study several commonly used detection statistics and analyze their performance under collusion attacks. We further propose a preprocessing technique of the extracted fingerprints specifically for collusion scenarios to improve the detection performance.

Algorithms↗

Fingerprint pattern restoration by digital image processing techniques.

Fingerprint evidence plays an important role in solving criminal problems. However, defective (lacking information needed for completeness) or contaminated (undesirable information included) fingerprint patterns make identifying and recognizing processes difficult. Unfortunately. this is the usual case. In the recognizing process (enhancement of patterns, or elimination of "false alarms" so that a fingerprint pattern can be searched in the Automated Fingerprint Identification System (AFIS)), chemical and physical techniques have been proposed to improve pattern legibility. In the identifying process, a fingerprint examiner can enhance contaminated (but not defective) fingerprint patterns under guidelines provided by the Scientific Working Group on Friction Ridge Analysis, Study and Technology (SWGFAST), the Scientific Working Group on Imaging Technology (SWGIT), and an AFIS working group within the National Institute of Justice. Recently, the image processing techniques have been successfully applied in forensic science. For example, we have applied image enhancement methods to improve the legibility of digital images such as fingerprints and vehicle plate numbers. In this paper, we propose a novel digital image restoration technique based on the AM (amplitude modulation)-FM (frequency modulation) reaction-diffusion method to restore defective or contaminated fingerprint patterns. This method shows its potential application to fingerprint pattern enhancement in the recognizing process (but not for the identifying process). Synthetic and real images are used to show the capability of the proposed method. The results of enhancing fingerprint patterns by the manual process and our method are evaluated and compared.

Dermatoglyphics↗

An evaluation of ERIC PCR and AP PCR fingerprinting for discriminating Salmonella serotypes.

PCR fingerprints of 89 Salmonella isolates belonging to 22 serotypes were obtained using ERIC PCR (enterobacterial repetitive intergenic consensus PCR) and AP PCR (arbitrarily primed PCR) to evaluate the ability of different fingerprinting methods to differentiate or identify serotypes and subtypes. Fingerprints were scored and comparisons were made using a computer program. ERIC PCR produced a unique, complex fingerprint for almost every isolate, but these fingerprints did not identify serotypes. One AP PCR primer also produced complex fingerprints that discriminated among isolates, but again did not identify serotypes. A second AP PCR primer produced simple patterns, including one pattern shared by 35 isolates from 12 different serotypes. In general, the three sets of PCR fingerprints distinguished isolates, but were not correlated with serotypes. Matching fingerprints from different gels by computer was difficult, since similarities were based on both intense and faint bands. In addition, this study suggests that dendrograms created from PCR fingerprints should be viewed with caution.

Base Sequence↗

Identifying the origin of single corneal cells by DNA fingerprinting: part II-- application to limbal allografting.

PURPOSE: Successful limbal allotransplantation allows regression of limbal stem cell deficiency features. Transplant survival is presumed if clinical improvement occurs. However, positive proof of surviving transplanted stem cells remains difficult. This follow-up study attempted to prove donor cell survival 5 years after limbal stem cell allograft in one woman with aniridia. METHODS: Impression cytology and single-cell DNA fingerprinting were used to investigate a previously studied patient. Corneal epithelial cells were harvested from five sites and isolated by micromanipulation. Polymerase chain reaction and short tandem repeat profiling were used to obtain forensic standard "DNA fingerprints" from single cells. (The technique is described in the preceding article, Part I.) Blood samples yielded host and donor DNA for comparison. Negative controls were performed for impression cytology and polymerase chain reaction. Simultaneous micro-scrape samples were also taken. RESULTS: Impression cytology samples permitted informative DNA fingerprints from all corneal sites and represented 76% (23/30) of tested cells. Fifty percent (15/30) of the fingerprints were "specific" but 83% (19/23) matched the host DNA fingerprint. The remaining 17% (4/23) represented contamination from various sources. Specific fingerprints were obtained in 55% (10/18) of the cells from micro-scrape samples. All samples giving sufficient information matched the host DNA fingerprint. All tested blood samples gave specific fingerprints. None of the sampled corneal cells gave a donor DNA fingerprint. CONCLUSIONS: In a single patient, no detectable long-term donor cell survival exists at 5 years. Positive identification would have provided unequivocal proof of donor cell survival. This technique gives useful information even if contamination occurs.

Aniridia↗