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At least 199 records · Page 11Linked to original sources

[Study on HPLC fingerprint of different growing areas on fructus aurantii fried with bran].

OBJECTIVE: To establish a method for HPLC fingerprint determination of different growing areas on Fructus Aurantii fried with bran. METHOD: HPLC chromatographic experiment was carried out by ultrasonic distilment of acetacetic ester, gradient elution mode and the detection wavelength of 320 nm. The column temperature was set at 30 degrees C, and flow rate is 1.0 mL x min(-1). Naringin served as the reference standard. Ten batches of Fructus Aurantii fried with bran in the different places were analyzed. Similarity evaluation system for chromatographic fingerprint of TCM proceeded the evaluation, The correlation coefficients between ten batches were used to define the similarity. RESULT: There were eleven characteristic peaks in Fructus Aurantii fried with bran by HPLC fingerprint which constituted fingerprint characters. Both the reproducibility and stability on HPLC fingerprint were proper. CONCLUSION: The method was accurate and simple. It can be applied to the analysis on Fructus Aurantii fried with bran. The fingerprint spectrum can be used to distinguish Fructus Aurantii fried with bran and to be its quality control.

China↗

[A chromatographic fingerprinting-based approach to evaluating the stability of production process of traditional Chinese medicine].

OBJECTIVE: To study a new method for the evaluation of the stability of production process of Traditional Chinese Medicines (TCM) based on chromatographic fingerprinting. METHOD: Taking the extraction process of Red Ginseng for example, the fingerprints of extracts produced with fluctuating processing factors, i.e. ratio of solvent to liquid, solvent concentration, extraction durations and extraction times, were acquired. Meanwhile, the similarity of fingerprinting based on Euclidean distance was calculated and each fingerprint was fitwith cubic spline. RESULT: Similarities of fingerprints of the extracts responded to the fluctuation of processing factors, and it showed that taking similarity as index, quantitative relation between chromatographic fingerprints and processing factors could be established. CONCLUSION: The method proposed in the paper was able to describe the quality of products resulting from fluctuating conditions in extracting process, and therefore, can be developed as a practicable strategy to evaluate the stability Traditional Chinese Medicines in production process.

Chromatography, High Pressure Liquid↗

Development of latent fingerprints using a corona discharge.

A novel technique for the development of latent fingerprints is presented. It is based on placing a fingerprint-bearing object inside a corona discharge induced plasma. The development of various real and artificial fingerprints on metallic substrates under a wide range of conditions is studied. Using the results of the development experiments and the results of X-ray photoelectron spectroscopy, it is shown that the development is based on oxidation of the fingerprint background. This is achieved by strong oxidizers generated by the discharge process, while saturated fatty-acids found in sebaceous fingerprints protect the area beneath them, resulting in a visible fingerprint. The process is optimized by minimizing the electrode gap distance and maximizing the peak discharge voltage and the pulse repetition frequency.

Dermatoglyphics↗

A study to evaluate the parent-offspring similarity in the maxillofacial profile using fingerprints in Japanese families.

An attempt was made to investigate the parent-offspring similarity in the maxillofacial profile by a quantitative analysis of fingerprints. Lateral roentgenographic cephalograms and ten fingerprints were obtained from 98 Japanese patients (44 males and 54 females) and their parents. Analysis showed a significant parent-offspring correlation for both maxillofacial profile and fingerprints. The relatively low father-son correlation for both features suggests a major influence of X-linked genes. The genetic correlation between the maxillofacial profile and fingerprints was significant for parent-son but not for parent-daughter pairings. The parent-offspring correlation in the maxillofacial profile was evaluated in two groups showing differences (distant group) or similarities (near group) in the fingerprint patterns between the father and mother. In the distant group, a greater parent-offspring similarity was observed in the maxillofacial profile. The same finding was also obtained on using only digit II of the parents. Therefore, from a morphogenetical point of view, parental fingerprints appear helpful in clinical applications designed to predict maxillofacial growth in offspring.

Adolescent↗

Calculating paternity probabilities from DNA multilocus fingerprints in some cases of deficiency.

Paternity determination via DNA multilocus fingerprints in normal trio cases, namely when the fingerprints of the trio mother, child and putative father are available, is performed on a routine basis. But in cases of deficiency, where the DNA fingerprint of mother or putative father is missing, there exists no systematic, analytical way for paternity calculations, even in cases where DNA fingerprints of other persons related to the missing individual are available. The aim of this paper is to establish a mathematical, analytical background, that enables paternity calculations in cases of deficiency if complementary information can be obtained from the DNA fingerprints of at least one of the parents of the missing individual. Using the basic outline of our previously described procedure, the formulas for paternity calculations also allow mutations.

Chromosome Mapping↗

Polymerase chain reaction fingerprinting in fungi using single primers specific to minisatellites and simple repetitive DNA sequences: strain variation in Cryptococcus neoformans.

Minisatellites and simple repetitive DNA sequence motifs are used as conventional oligonucleotide probes in DNA-hybridization-based fingerprinting. The same oligonucleotides can be used as single primers in the polymerase chain reaction (PCR) to generate individual PCR fingerprints. In this study, the simple repetitive sequences, (CA)8, (CT)8, (CAC)5, (GTG)5, (GACA)4 and (GATA)4, and a minisatellite core sequence derived from the wild-type phage M13 (5' GAGGGTGGCGGTTCT 3') were used as specific, single primers to amplify hypervariable repetitive DNA sequences during PCR analysis. The potential applications of this techniques are demonstrated with clinical isolates of the human pathogenic yeast, Cryptococcus neoformans. PCR fingerprint patterns have remained stable after long-term in vitro passage ( > 2 1/2 years to date). Hybridization of the primers to blots of electrophorectically separated chromosomes demonstrated that the target sequences recognized by most of the primers are dispersed through the entire yeast genome. Sequence analysis of the cloned bands obtained by PCR fingerprinting indicated that if the same or extremely similar, inversely oriented tandem repeats are located close to each other, when only one repeat-specific primer is used in the PCR, the region between these repeats is amplified. PCR fingerprinting has a wide range of current and potential applications to fungi, such as clarifying taxonomic questions, facilitating epidemiological studies and improving the diagnosis of mycotic diseases.

Base Sequence↗

Application of DNA fingerprinting to the taxonomy of European blackberry species.

Thousands of blackberry species (Rubus subgenus Rubus) have been described from Central and Northern Europe. These species are usually polyploid and pseudogamous. Since apomixis is only facultative, the species have retained the possibility to produce some offspring by sexual recombination. Hybridization of restriction-enzyme-digested DNA samples to the M13, (AC)/(TG) and 33.15 probes has yield valuable information on the occurrence and distribution of genetic variation in Rubus. Intraspecific variation proved to be very restricted, with identical DNA fingerprints sometimes being exhibited by Swedish, Danish and German populations of the same species. By comparison, interspecific variation was usually pronounced, suggesting that most species have been derived through genetic recombination. Speciation through interspecific hybridization was demonstrated in one case. Several critical taxa were investigated; some of these proved to have unique DNA fingerprints, whereas others appear to be identical with previously described species. Evaluation of raspberry cultivars, obtained through several generations of vegetative propagation or with various in vitro techniques, failed to detect any DNA fingerprint variation, suggesting that the fingerprints are somatically stable. Blackberry seedlings, obtained by pollinating various species as well as interspecific hybrids with pollen from yet other species, were successfully evaluated with DNA fingerprinting to determine the relative proportions of the apomictic and sexual seed set.

DNA Fingerprinting↗

Extra-pair sires as identified by means of standardized across-gel comparisons of multilocus DNA fingerprints.

Molecular analyses of genetic parentage in wild bird populations demonstrate that extra-pair paternity (EPP) is common, even in socially monogamous species. After having identified extra-pair offspring (EPO), an important step is to uncover the identity of extra-pair sires (EPS). Studies relying on multilocus DNA fingerprinting face the problem that simple between-gel comparisons of fingerprint banding patterns are imprecise in assigning parentage across gels. To tackle this problem, we developed a method to identify EPS by means of standardized across-gel comparisons of multilocus DNA fingerprints and applied it in a socially monogamous bird species with high levels of EPP, the coal tit (Parus ater). Scanned DNA fingerprint gels and standard image-editing software allowed to screen and reliably identify EPS. Overall, EPP was successfully assigned to 32 different EPS for 143 out of 210 (68.1%) EPO detected. This corresponds well to extra-pair parentage assignment rates revealed by microsatellite-based studies in congeneric species. Our approach proves especially useful whenever multilocus DNA fingerprints for parentage exclusion exist while microsatellite markers are not available.

Animals↗

'Fingerprinting' of HLA-DQA by polymerase chain reaction and heteroduplex analysis.

We have developed a rapid, non-radioisotopic PCR fingerprinting technique for analysis of the HLA-Class II DQA gene second exon polymorphism, and have applied it to DNA samples from 210 healthy individuals. The technique is based on the formation of specific patterns (fingerprints) of homoduplexes or heteroduplexes between in vitro amplified DNA sequences. After electrophoresis on non-denaturing polyacrylamide gels and ethidium bromide fluorescence or silver staining, different HLA-DQA types give allele-specific banding patterns. HLA DQA typing is done by visual comparison between the sample's fingerprint patterns and appropriate controls. Similar fingerprints can be resolved by mixing the sample with a standard DNA in an amplified 'DNA crossmatch'. This application of PCR fingerprinting is useful to confirm the HLA-DQA serological typing and to improve the molecular characterization of this polymorphic region.

Alleles↗

Oligonucleotide fingerprinting using simple repeat motifs: a convenient, ubiquitously applicable method to detect hypervariability for multiple purposes.

A panel of simple repetitive oligonucleotide probes has been designed and tested for multilocus DNA fingerprinting in some 200 fungal, plant and animal species as well as man. To date at least one of the probes has been found to be informative in each species. The human genome, however, has been the major target of many fingerprinting studies. Using the probe (CAC)5 or (GTG)5, individualization of all humans is possible except for monozygotic twins. Paternity analyses are now performed on a routine basis by the use of multilocus fingerprints, including also cases of deficiency, i.e. where one of the parents is not available for analysis. In forensic science stain analysis is feasible in all tissue remains containing nucleated cells. Depending on the degree of DNA degradation a variety of oligonucleotides are informative, and they have been proven useful in actual case work. Advantages in comparison to other methods including enzymatic DNA amplification techniques (PCR) are evident. Fingerprint patterns of tumors may be changed due to the gain or loss of chromosomes and/or intrachromosomal deletion and amplification events. Locus-specific probes were isolated from the human (CAC)5/(GTG)5 fingerprint with a varying degree of informativeness (monomorphic versus truly hypervariable markers). The feasibility of three different approaches for the isolation of hypervariable mono-locus probes was evaluated. Finally, one particular mixed simple (gt)n(ga)m repeat locus in the second intron of the HLA-DRB genes has been scrutinized to allow comparison of the extent of exon-encoded (protein-) polymorphisms versus intronic hypervariability of simple repeats: adjacent to a single gene sequence (e.g. HLA-DRB1*0401) many different length alleles were found. Group-specific structures of basic repeats were identified within the evolutionarily related DRB alleles. As a further application it is suggested here that due to the ubiquitous interspersion of their targets, short probes for simple repeat sequences are especially useful tools for ordering genomic cosmid, yeast artificial chromosome and phage banks.

Animals↗

DNA fingerprinting of trait-selected mouse lines and linkage analysis in reference families.

The first aim of the study was the molecular genetic characterization of long-term trait-selected (growth, adaptability and fertility) lines of mice using multilocus DNA fingerprinting with the simple tandem repetitive oligonucleotide probes (GAA)6 and (GACA)4. Secondly polymorphic markers were screened for association with growth performance based on DNA fingerprint analysis in reference families. Pooled DNA samples of ten unrelated mice of trait-selected mouse lines (over 40 generations) and the unselected control lines were analyzed. Resulting differences in band patterns were reanalyzed comparing individual fingerprints of the animals included in the pooled DNA samples of the different lines. Between the mouse lines about 30% of analyzed fingerprint bands were polymorphic. Individuals of long-term selected mouse lines show only a few individual-specific bands. Most polymorphic bands observed in DNA mixes appear in all animals included in the DNA mix of the corresponding line. Line-specific DNA fingerprint bands were analyzed for their inheritance and linkage with growth performance in reference families using animals with extreme growth performance of the first backcross after crossing of growth-selected with unselected mice. Scanning the distribution of line-specific bands in the reference panel few bands were identified which are associated with growth performance. They appear as useful markers for growth selection. Nevertheless most line-specific bands result from genetic drift rather than from selection.

Adaptation, Physiological↗

Pulsed-field gel electrophoresis fingerprinting for identification of Azospirillum species.

Pulsed-field gel electrophoresis (PFGE) was used to obtain macrorestriction fingerprints of restriction enzyme-cut DNA of natural isolates of Azospirillum spp. Metabolic profiles, along with other phenotypic characteristics, were compared with these fingerprints to differentiate among the azospirilla isolates. A wide diversity of phenotypes (e.g., colony color, motility, and accumulation of poly-beta-hydroxybutyrate granules) was observed among the natural isolates of azospirilla. PFGE revealed that TCTAGA, the sequence recognized by Xba1, is rare in the genome of azospirilla. The PFGE fingerprint revealed that azospirilla associated with different crops have a very similar genetic background. PFGE fingerprints were more consistent in the identification of azospirilla isolates from specific hosts than the metabolic fingerprints. For further differentiation at strain level, metabolic, physiological, and morphological profiles provide additional information.

Azospirillum↗

Fingerprinting reveals gamma-ray induced mutations in fungal DNA: implications for identification of patent strains of Trichoderma harzianum.

We have analyzed different patent strains and gamma-ray induced mutants of Trichoderma harzianum by DNA fingerprinting and PCR fingerprinting (RAPD). Applying wild-type phage M13 DNA, with the oligonucleotides (CT)8 and (GTG)5 as probes for hybridization, as well as the oligonucleotides GGCATCGGCC, (GTG)5, (CAC)5 and the M13 sequence GAGGGTGGCGGTTCT as primers in PCR, we were able to obtain different and discriminative fingerprint patterns for all strains and mutants investigated. Irradiation of fungi led to mutations which resulted in new fingerprint patterns. Consequently, irradiation-induced mutants can be clearly distinguished from the original wild-type isolates by genomic fingerprinting which is of importance for the patent protection of fungal strains. Sequencing of the ITS-1 and ITS-2 regions of the rDNA gene complex revealed the same sequence for all mutant strains and the original wild-type strain.

Base Sequence↗

Heterogeneity of Escherichia coli derived from artiodactyla animals analyzed with the use of rep-PCR fingerprinting.

Genetic polymorphism of 83 isolates of E. coli, derived from 4 species of artiodactyla animals living in a relatively close contact on the grounds of a theme park ZOO Safarii Swierkocin (Poland) was determined using the rep-PCR fingerprinting method, which utilizes oligonucleotide primers matching interspersed repetitive DNA sequences in PCR reaction to yield DNA fingerprints of individual bacterial isolates based on repetitive extragenic palindrome (REP) primers. The fingerprint patterns demonstrated the essential polymorphism of distribution of REP sequences in genomes of the examined isolates. The arithmetic averages clustering algorithm (UPGMA) statistical analysis of fingerprints with the use of the Jaccard similarity coefficient differentiated E. coli isolates into three similarity groups containing various numbers of isolates. The groups comprised isolates derived from two, three and four species of the source animals. The isolates derived from each source segregated in the dendrogram in a different way, both within the similarity groups and among them, indicating an individual repertoire of E. coli in the examined species of animals. The similarity relations among E. coli derived from the same source, illustrated in a dendrogram with a number of subclusters of a low mutual similarity (< or = 20%), indicated an essential interstrain differentiation in terms of the distribution of REP sequences. Our results confirmed the hypothesis of the oligoclonal characters of populations obtained from particular sources. The rep-PCR fingerprinting method with REP primers is simple and highly differentiating and can be recommended for use in explorations of large groups of animals and monitoring the variability of strains.

Animals↗

Improved strategy for comparing microbial assemblage fingerprints.

Microbial fingerprinting techniques permit the rapid visualization of entire assemblages in single assays, allowing direct comparison of communities in different samples, where the null hypothesis of such analyses is that all samples are the same. The comparison of fingerprints relies upon the precise estimation of all amplified DNA fragment lengths, which correspond to operational taxonomic units (OTU; analogous, but not equal to, a taxon in macroorganism studies). However, computer interpolation of size standards (and consequently OTU size calling) can be imprecise between gel runs, which can lead to imprecise calculation of similarity indices between multiple assemblages. To account for OTU size calling imprecision, all fragments within a range of sizes (a window) can be combined (i.e., "binned") where the window is as wide as the imprecision of OTU size calling. However, artifacts may occur upon binning samples that may cause samples to appear less similar to each other, caused by splitting of OTU between adjacent bin windows. In this work we present an improved binning technique that accounts for OTU size calling imprecision in the comparison of multiple fingerprints. This technique comprises binning all pairwise comparisons in multiple bin window frames, where the starting size of the window (i.e., frame) is shifted by +1 bp for a total of x frames, where x bp is the width of the maximum bin window size in any binning scheme. Pairwise similarity indices between different community fingerprints are calculated for each of the x frames. To best address the null hypothesis of the community comparison, the maximum similarity value of all x frames is then used in downstream analyses to compare the communities. We believe this binning technique provides the most accurate and least biased comparison between different microbial fingerprints.

Artifacts↗

Protein extraction and fingerprinting optimization of bacterial communities in natural environment.

Recent development in molecular approaches allows access to genetic structure and diversity of indigenous microbial communities. In contrast, the functional analysis of microorganisms in their environment is still hampered by methodological limitations. Analysis of total proteins expressed at the whole community level (metaproteome) has been proposed to characterize the functional structure of microbial communities in their environment. However, developments are still required to perform such analysis. Our aim was to optimize methods to extract and characterize metaproteome of indigenous microbial communities. Experiments were first conducted in monoxenic bacterial cultures, and various methods were examined to define a procedure of protein extraction ensuring an efficient recovery regardless of the taxonomic affiliation of the cells. These developments were next applied to characterize the metaproteome from indigenous bacterial communities in freshwater samples. Bacterial cells were recovered from water using a high-speed density gradient centrifugation method before protein extraction and fingerprinting. The reactivity and sensitivity of this metaproteomic approach were tested by analyzing the variations of protein fingerprints according to perturbations (cadmium or mercury contamination). The genetic structure of the corresponding communities was also characterized by automated ribosomal spacer analysis (ARISA) DNA fingerprinting. Both protein and DNA fingerprints were statistically analyzed. Results obtained showed that the method developed for protein recovery and fingerprinting was efficient, sensitive, and reproducible. Both the functional and genetic structures of the freshwater bacterial community were complex and varied with perturbations. These variations occurred at both population and protein expression levels and were specific to the perturbation applied.

Bacteria↗

PCR-fingerprinting for selection of HLA matched unrelated marrow donors. Collaborating Centres in the IMUST Study.

HLA typing contributes to the delays that occur in the search for HLA-matched unrelated marrow donors, and that result in poor patient survival. A new DNA technique for testing DR match between patient and unrelated marrow donors has been assessed. The technique is based on the formation of heteroduplexes between heterologous amplified coding and non-coding DNA sequences during the final annealing stage of the polymerase chain reaction (PCR), and different HLA-DR/Dw types give unique banding patterns ("PCR fingerprints") on non-denaturing polyacrylamide gel electrophoresis. HLA-DR matching is by visual comparison of patients' with donors' fingerprints. Identity can be confirmed by mixing donor and recipient DNA before the final stage of the PCR ("DNA crossmatching"). In an assessment of the technique in 53 unrelated HLA-A and HLA-B matched patient-donor pairs, 42 pairs gave the same results with PCR fingerprinting as with DNA-RFLP analysis. In the 11 other pairs DR/Dw mismatches were detected by PCR fingerprinting but not by the standard DNA-RFLP method; PCR-SSO typing with selected sequence-specific oligonucleotides (SSO) confirmed that mismatches were due to different subtypes of DR4. PCR fingerprinting might thus accelerate the selection of unrelated marrow donors by simplifying the logistics of the donor search.

DNA Fingerprinting↗

Restriction polymorphisms and fingerprint patterns from an interspersed repetitive element of Plasmodium falciparum DNA.

A recombinant DNA clone, pC4.H32, identifies distinguishable restriction fragment patterns from different Plasmodium falciparum clones. Analysis of these DNA fingerprint patterns from parasites cultivated over several years and from progeny of a P. falciparum cross showed the fingerprints to be mitotically and meiotically stable. Restriction fragments from the parents of the cross possessed sufficient polymorphism and number to generate 14 unique fingerprint patterns in 16 independent recombinant progeny. The pC4.H32 insert contains a 0.5-kb imperfectly repeated sequence found in subtelomeric regions of multiple chromosomes. Restriction site variations both within and outside of the 0.5-kb repeat contribute to the fingerprint polymorphisms. Fingerprint analysis can serve to type P. falciparum clones and can detect mislabeling and cross-contamination of parasite stocks.

Animals↗