Genetic variation in Przewalski's horses, with special focus on the last wild caught mare, 231 Orlitza III.

In our continuing efforts to document genetic diversity in Przewalski's horses and relatedness with domestic horses, we report genetic variation at 22 loci of blood group and protein polymorphisms and 29 loci of DNA (microsatellite) polymorphisms. The loci have been assigned by linkage or synteny mapping to 20 autosomes and the X chromosome of the domestic horse (plus four loci unassigned to a chromosome). With cumulative data from tests of 568 Przewalski's horses using blood, hair or tooth samples, no species-defining markers were identified, however a few markers were present in the wild species but not in domestic horses. Inheritance patterns and linkage relationships reported in domestic horses appeared to be conserved in Przewalski's horses. A derived type for the last wild caught mare 231 Orlitza III provided evidence for markers apparently not found in (or not currently available by descent from) the other species founders that were captured at the end of the nineteenth century. This information has been critical to the development of parentage analyses in the studbook population of Przewalski's horses at Askania Nova, at one time the largest herd of captive animals and the source of stock for reintroduction efforts. Some horses in the study showed genetic incompatibilities with their sire or dam, contradicting published studbook information. In many cases alternative parentage could be assigned from living animals. To assist in identification of correct parentage, DNA marker types for deceased horses were established from archived materials (teeth) or derived from offspring. Genetic markers were present in pedigreed animals whose origin could not be accounted for from founders. Genetic distance analysis of erythrocyte protein, electrophoretic and microsatellite markers in Przewlaski's horses and ten breeds of domestic horse place the Przewalski's horse as an outgroup to domestic horses, introgression events from domestic horses not withstanding.

Proton transverse nuclear magnetic relaxation in oxidized blood: a numerical approach.

When red blood cells are deoxygenated, hemoglobin, which is then transformed into deoxyhemoglobin or methemoglobin, becomes paramagnetic. The transverse nuclear magnetic relaxation rate of water protons is considerably enhanced by this chemical transformation. A general agreement exists about the origin of the phenomenon--local field inhomogeneities induced by paramagnetic centers randomly distributed within the cell--but the localization of the region that dominates the relaxation is unclear. We addressed this problem with a computer simulation devoted to the determination of transverse magnetic relaxation of water protons in the presence of superparamagnetic MRI contrast agent candidates. The simulation confirms an earlier experimental result that shares equitably the responsibility for the observed relaxation between intracellular and extracellular water.

Structure and dynamics of water in tendon from NMR relaxation measurements.

Nuclear magnetic relaxation times were measured in collagen tissue when varying the orientation of the fiber with respect to the static field. T1 was found to be only slightly dependent on theta, the fiber-to-field angle, but T2 was very sensitive to the orientation, with a maximum value at the magic angle. The transverse decay curves were multiexponential. Their deconvolution displayed four components; the ones that decayed most slowly were almost independent of theta, but the two fastest ones showed a strong angular dependence that was interpreted with a cross-relaxation model. Quadrupolar dips were visible in the 1/T1 dispersion curves. These dips were independent of theta, so that the magnetization transfer could also be assumed to be independent of the fiber orientation. Finally, each component was assigned to a fraction of protons localized in the macromolecular structure and characterized by particular dynamics. The model of Woessner was applied to the water molecules tightly bound into the macromolecules, which resulted in a dynamical description of this water fraction. This description is compatible with the two-sites model of Ramachandran based on x-ray diffraction and with the extensive studies of Berendsen. However, the important indications obtained from the deconvolution lead to a less static representation of the tissue.

Fiber-to-field angle dependence of proton nuclear magnetic relaxation in collagen.

Longitudinal and transverse proton relaxation times were measured on pig tendon. For T1, dispersion curves and more accurate measurements at 20 MHz are presented. Values of T2 were obtained from CPMG pulse sequences, at 20 MHz. The dependence of relaxation times against the fiber-to-field angle was particularly investigated. Longitudinal relaxation rate was found to be almost orientation independent, and presented quadrupolar peaks between 1 and 4 MHz. On the contrary, transverse relaxation, that was well fitted by the sum of four exponentials, was highly orientation dependent. Deconvolution showed that the exponentials decaying most quickly are most orientation dependent. For those two fractions, a cross-relaxation model allowed explaining the fiber-to-field angle dependence, and the specially low rate corresponding to the magic angle of 55 degrees. Finally, each decaying mode was assigned to a fraction of protons localized in the macromolecular structure and characterized by particular dynamics.