ABSTRACT
Coat colour variation is determined by many genes, one of which is the melanocortin receptor type 1 (MC1R) gene. In this study, we examined the whole coding sequence of this gene in four species belonging to the Canidae family (dog, red fox, arctic fox and Chinese raccoon dog). Although the comparative analysis of the obtained nucleotide sequences revealed a high conservation, which varied between 97.9 and 99.1%, we altogether identified 22 SNPs (10 in dogs, six in farmed red foxes, two in wild red foxes, three in arctic foxes and one in Chinese raccoon dog). Among them, seven appeared to be novel: one silent in the dog, three missense and one silent in the red fox, one in the 3'-flanking region in the arctic fox and one silent in the Chinese raccoon dog. In dogs and red foxes, the SNPs segregated as 10 and four haplotypes, respectively. Taking into consideration the published reports and results of this study, the highest number of missense polymorphisms was until now found in the dog (9) and red fox (7).
Subject(s)
Dogs/genetics , Foxes/genetics , Polymorphism, Genetic , Raccoon Dogs/genetics , Receptor, Melanocortin, Type 1/genetics , Animals , Hair , Pigments, Biological/geneticsABSTRACT
BACKGROUND: Mycobacterium tuberculosis is one of the most dangerous human pathogens. Molecular typing of M. tuberculosis has allowed better control of tuberculosis and, among other benefits, identification of genetic lineages among strains. OBJECTIVE: To test the potential of polymerase chain reaction (PCR) based methods for the epidemiological study of M. tuberculosis strains isolated from patients residing in a single city. DESIGN: We performed spoligotyping, mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) typing and insertion sequence (IS) 6110 restriction fragment length polymorphism (RFLP) analyses of 234 clinical strains of M. tuberculosis collected over 2 years from the Polish city of Lodz. RESULTS: Spoligotyping analysis revealed 84 spoligotypes with a shared international type and 50 unique spoligotypes. Subtyping via 15- and 19-loci MIRU-VNTR analyses revealed 154 patterns with 117 unique profiles, and 159 patterns with 126 unique profiles, respectively. Spoligotyping combined with MIRU-VNTR 15- and 19 loci analyses revealed 132 and 146 unique profiles, respectively. Overall, 96 strains clustered via MIRU-VNTR typing were used in IS6110-RFLP analysis. Complete congruence of patterns revealed by PCR-based methods was noted for 40 strains, of which 36 were isolated from epidemiologically linked patients. CONCLUSION: The combination of 15-loci MIRU-VNTR typing with spoligotyping is useful for primary analysis of M. tuberculosis strains; however, additional use of MIRU 23 should be considered. Strains clustered by PCR-based methods should be further analysed by IS6110-RFLP typing.
Subject(s)
Molecular Typing/methods , Mycobacterium tuberculosis/isolation & purification , Polymorphism, Restriction Fragment Length , Tuberculosis/microbiology , Adolescent , Adult , Aged , Bacterial Typing Techniques/methods , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Molecular Epidemiology/methods , Mycobacterium tuberculosis/genetics , Poland/epidemiology , Polymerase Chain Reaction/methods , Tandem Repeat Sequences , Tuberculosis/epidemiology , Young AdultABSTRACT
The aim of this paper was to evaluate the usefulness of digital image analysis techniques to measure the amount and strength of immunohistochemical markers. The new method, based on the spatial visualization technique, was confronted with methods of colour sampling and grey scale thresholding. Examples of applications of the techniques for apoptosis and proliferation markers are also presented.
Subject(s)
Goiter/pathology , Immunohistochemistry/methods , Biomarkers/analysis , Humans , Image Processing, Computer-Assisted , Ki-67 Antigen/analysis , Proliferating Cell Nuclear Antigen/analysisABSTRACT
The aim of this study was an application of spatial visualization techniques for quantitative measurements of immuno- and histochemical reactions. For a quantitative histochemical study, specimens, collected from patients with chronic gastritis, were stained with paS/AB, while for immunohistochemical evaluation, specimens were used, collected from patients with chronic parathyroiditis and were analyzed with Ki-67 proliferation marker and apoptosis bcl-2 protein. The new technique permitted to obtain quantitative objective results. Statistical cluster analysis of those results indicated small groups of cases for reevaluation and supported the final diagnosis.
