Polymorphism analysis of four canine MHC class I genes

Abstract: We have studied the variability of four structurally complete dog leukocyte antigen (DLA) class I genes, termed DLA-12, -88, -79 and -64, in a population of mixed breed, unrelated dogs. The human HLA and canine DLA loci share a high degree of similarity in terms of gene structure. This analysis focused on the first three exons of each of four complete canine genes. Exons two and three are the major source of polymorphism in the corresponding human genes. In this analysis, DLA-88 was found to be significantly more polymorphic than the other three genes, with 44 distinct alleles observed among 63 mixed breed, unrelated dogs. The remaining genes had between one and four alleles when examined in 25 dogs. This work was carried out as part of an effort to develop an MHC typing system for the dog, which is critical to the further development of preclinical studies of hematopoietic stem cell and solid organ transplantation in the canine model.     

Localisation of myocardial ischaemia from the magnetocardiogram using current density reconstruction method: computer simulation study

A computer simulation study is performed to investigate the method of current density reconstruction to localise myocardial ischaemia. A computer model of the entire human heart is used to simulate the excitation and repolarisation process in eight topographically different cases of myocardial ischaemia. The associated magnetocardiogram is calculated at 37 positions of the KRENIKON^* biomagnetic measurement equipment. The method of current density reconstruction is applied at the S-point (the last discemible deviation from the ST-segment at the end of the QRS-complex) of the MCG to find characteristics of the myocardial ischaemia simulated by the model. The results show that it is possible to determine the location of the ischaemia. The current density distribution may be interpreted physiologically in terms of the so-called ‘injury current’. This indicates that magnetocardiography might be a suitable method for noninvasive ischaemia diagnosis, and further investigations of the current density reconstruction method for the injury current should be performed on patients with ischaemic heart disease.     

Dynamics of the bacterial SMC complex and SMC-like proteins involved in DNA repair

Bacteria and archaea possess several different SMC-like proteins, which perform essential functions in a variety of chromosome dynamics, such as chromosome compaction, segregation, and DNA repair. SMC-like proteins localize to distinct sites within the cells at different time points in the cell cycle, or are recruited to sites of DNA breaks and damage. The bacterial SMC (MukB) complex appears to perform a condensin-like function, while SbcC and RecN act early during DNA repair, but apparently at different sites within the cells. Thus, bacterial SMC-like proteins have dynamic functions in chromosome segregation and maintenance of genetic stability.     

Aminochrome as a preclinical experimental model to study degeneration of dopaminergic neurons in Parkinson’s disease

Four decades after L-dopa introduction to PD therapy, the cause of Parkinson's disease (PD) remains unknown despite the intensive research and the discovery of a number of gene mutations and deletions in the pathogenesis of familial PD. Different model neurotoxins have been used as preclinical experimental models to study the neurodegenerative process in PD, such as 6-hydroxydopamine (6-OHDA), 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), and rotenone. The lack of success in identifying the molecular mechanism for the degenerative process in PD opens the question whether the current preclinical experimental models are suitable to understand the degeneration of neuromelanin-containing dopaminergic neurons in PD. We propose aminochrome as a model neurotoxin to study the neurodegenerative processes occurring in neuromelanin-containing dopaminergic neurons in PD. Aminochrome is an endogenous compound formed during dopamine oxidation and it is the precursor of neuromelanin, a substance whose formation is a normal process in mesencephalic dopaminergic neurons. However, aminochrome itself can induce neurotoxicity under certain aberrant conditions such as (i) one-electron reduction of aminochrome catalyzed by flavoenzymes to leukoaminochrome o-semiquinone radical, which is a highly reactive neurotoxin; or (ii) the formation of aminochrome adducts with alpha-synuclein, enhancing and stabilizing the formation of neurotoxic protofibrils. These two neurotoxic pathways of aminochrome are prevented by DT-diaphorase, an enzyme that effectively reduces aminochrome with two-electrons preventing both aminochrome one-electron reduction or formation alpha synuclein protofibrils. We propose to use aminochrome as a preclinical experimental model to study the neurodegenerative process of neuromelanin containing dopaminergic neurons in PD.     

Bacterial translation elongation factor EF-Tu interacts and colocalizes with actin-like MreB protein

We show that translation initiation factor EF-Tu plays a second important role in cell shape maintenance in the bacterium Bacillus subtilis. EF-Tu localizes in a helical pattern underneath the cell membrane and colocalizes with MreB, an actin-like cytoskeletal element setting up rod cell shape. The localization of MreB and of EF-Tu is interdependent, but in contrast to the dynamic MreB filaments, EF-Tu structures are more static and may serve as tracks for MreB filaments. In agreement with this idea, EF-Tu and MreB interact in vivo and in vitro. Lowering of the EF-Tu levels had a minor effect on translation but a strong effect on cell shape and on the localization of MreB, and blocking of the function of EF-Tu in translation did not interfere with the localization of MreB, showing that, directly or indirectly, EF-Tu affects the cytoskeletal MreB structure and thus serves two important functions in a bacterium.     

Multiple-spin-echo imaging with a 2D Fourier method

In 2D Fourier imaging the normal Carr-Purcell multiple-echo sequence generally leads to center line and mirror artifacts caused by imperfect rotations by the rf pulses. We describe a method to avoid these distortions using a phase alternating-phase shift (PHAPS) sequence which also allows multiple-slice and multiple-echo imaging at the same time. Measuring phantoms with calibrated T2 values, we have shown that the PHAPS imaging sequence leads to an accuracy of quantitative T2 determinations of better than 10%. Contrast-enhanced images are presented which we calculated from multiple-echo images and extrapolated to arbitrary echotimes, including negative ones. We believe that these improvements in T2 imaging will result in a significant reduction of patient investigation time in magnetic resonance imaging.     

3D soft tissue imaging with a mobile C-arm.

We introduce a clinical prototype for 3D soft tissue imaging to support surgical or interventional procedures based on a mobile C-arm. An overview of required methods and materials is followed by first clinical images of animals and human patients including dosimetry. The mobility and flexibility of 3D C-arms gives free access to the patient and therefore avoids relocation of the patient between imaging and surgical intervention. Image fusion with diagnostic data (MRI, CT, PET) is demonstrated and promising applications for brachytherapy, RFTT and others are discussed.