Immunology

A precise organization of contractile proteins is essential for contraction of heart muscle. Without a necessary stoichiometry of proteins, beating is not possible. Disruption of this organization can be seen in diseases such as familial hypertrophic cardiomyopathy and also in acquired diseases. In addition, isoform diversity may affect contractile properties in such functional adaptations as cardiac hypertrophy. The Mexican axolotl provides an uncommon model in which to examine specific proteins involved with myofibril formation in the heart. Cardiac mutant embryos lack organized myofibrils and have altered expression of contractile proteins. In order to replicate the disruption of myofibril formation seen in mutant hearts, we have developed procedures for the introduction of contractile protein antibodies into normal hearts. Oligonucleotides specific to axolotl tropomyosin isoforms (ATmC-1 and ATmC-3), were also successfully introduced into the normal hearts. The antisense ATmC-3 oligonucleotide disrupted myofibril formation and beating, while the sense strands did not. A fluoresceintagged sense oligonucleotide clearly showed that the oligonucleotide is introduced within the cells of the intact hearts. In contrast, ATmC-1 anti-sense oligonucleotide did not cause a disruption of the myofibrillar organization. Specifically, tropomyosin expression can be disrupted in normal hearts with a lack of organized myofi-brils. In a broader approach, these procedures for whole hearts are important for studying myofibril formation in normal hearts at the DNA, RNA, and/or protein levels and can complement the studies of the cardiac mutant phenotype. All of these tools taken together present a powerful approach to the elucidation of myofibrillogenesis and show that embryonic heart cells can incorporate a wide variety of molecules with cationic liposomes.

Aim. Anti-donor antibodies, denoted as "panel-reactive antibodies" (PRAs), are one of the most important factors influencing graft survival after renal transplantation. PRA is generally analyzed with enzyme-linked immunosorbent assay or flow cytometry (FC), which identify the HLA antigen specific for the preformed antibody. Patients and methods. We tested 66 patients for FC crossmatch (FCXM) when they were called for cadaveric renal transplantation. Thirty of 66 patients were FCXM-positive; 36 were FCXM-negative. Among the FCXM positive crossmatches, 21 were T-and B-cell positive; seven B positive; and two T positive. The HLA antibodies in the sera of FCXM-positive patients were reanalyzed using flow-PRA. Results. We detected HLA antibodies in 28/66 sera with flow PRA. The sera of 16/21 T-/B-, FCXM-positive patients contained both class I and II anti-HLA antibodies, five had only class I anti-HLA antibodies. One out of seven B-cell FCXM-positive patients had class I and class II anti-HLA antibodies, three, class I and 1 class II anti-HLA antibodies; the other two were negative. Class I and class II HLA antibodies were observed in two T-cell FCXM-positive patients. Four of 36 patients who were FCXM-negative were flow PRA positive: one had both class I and class II HLA antibodies and three, only class I HLA antibody. The comparison of FCXM and flow PRA results was significant (P ϭ .001). Conclusion. FCXM results may be confirmed by flow PRA tests, an important method to differentiate HLA versus non-HLA antibodies.

Objective. The aim of this study was to investigate the quality of life of renal donors during long-term follow-up. Patients and Methods. The short form health survey (SF-36) questionnaire was compared between renal donors and the general population. We evaluated the relationship to postoperative complications and preoperative information with the quality of life. Results. Fifty renal donors of mean age 55.8 Ϯ 12 years (range, 29 -70 years) had a mean follow-up of 55.1 Ϯ 47.2 months (range, 12-168 months). Complications after donor nephrectomy were related with physical function loss (r ϭ Ϫ.397; P Ͻ .05) and vitality (r ϭ Ϫ.463; P ϭ .01). Renal donor candidates who did not have satisfactory information before the operation experienced difficulty with decision making (r ϭ Ϫ.555; P ϭ .0001). Physical function, limitation of physical role and limitation of emotional role were comparable to the general population. Pain scale was worse among donors compared with the general population (P ϭ .001). Educational status of renal donors was related to the pain scale and vitality (r ϭ .369; P Ͻ .05 and r ϭ .523; P Ͻ .05, respectively). General health perception, vitality, mental health, and social functioning were worse compared with the general population (P ϭ .0001, P ϭ .002, P ϭ .0001, and P ϭ .001, respectively). Health problems occurring after donation were related to negation of interfamily relations (r ϭ .695; P ϭ .0001).

Widespread external and internal changes in body morphology have long been known to be hallmarks of the process of metamorphosis. However, more subtle changes, particularly at the molecular level, are only now beginning to be understood. A number of transcription factors have recently been shown to alter expression either in levels of message or in isoforms expressed. In this article, we describe a dramatic increase in the expression of the homeobox gene HoxA5 in the heart and aorta of the Mexican axolotl Ambystoma mexicanum during the process of thyroxin-induced metamorphosis. Immunohistochemical analysis with anti-HoxA5 antibody in thyroxin-induced metamorphosing animals showed a pattern of expression of HoxA5 comparable to that in spontaneously metamorphosing animals. Further, by in situ hybridization, we were able to show significant qualitative differences in the expression of this gene within the heart. Maximum HoxA5 expression occurred at the midpoint of metamorphosis in the myocardium, whereas the hearts of completely metamorphosed animals had the highest levels of expression in the epicardium and endocardium. In the aorta, smooth-muscle cells of the tunica media as well as cells of the tunica adventitia had an increase in expression of HoxA5 with thyroxin-induced metamorphosis. HoxA5 expression significantly changed in cells of the aorta and ventricle with treatment by thyroid hormone. HoxA5, a positive regulator of p53, may be involved with the apoptotic pathway in heart remodeling during amphibian metamorphosis.

The continuous GPS network operating on Mt. Etna with its 36 stations is currently one of the largest worldwide. The aim of this network is the evaluation of volcanic hazard and the modelling of the active sources. In this paper, we propose an in-depth analysis and modelling of continuous GPS data collected at Mt. Etna from May 2008 to December 2010. The analyzed period has been divided into four different coherent phases: 1) 14 May 2008-02 August 2008 (deflation of the entire GPS network); 2) 02 August 2008-14 June 2009 (deflation of the summit area and inflation at lower heights); 3) 14 June 2009-21 May 2010 (inflation of the entire GPS network); 4) 21 May 2010-31 December 2010 (inflation at medium and low heights and end of the inflation in the summit area). Analytical models indicate a non-uniform deformation style revealing spaced sources acting at different time on different segments of a multi-level magma reservoir. The Etnean plumbing system imaged here is depicted as an elongated magma reservoir that extends from the volcano body downwards to about 6.5 km below sea level (b.s.l.), sloping slightly towards the NorthWest , with storage volumes located at about 6.5, 2.0 and 0.0 km (b.s.l.). The changes in position of the modelled pressure sources during the analyzed time intervals indicate that, throughout the 2008 eruptive period, the deformation field was mostly driven by the upward migration of magma. On the other hand, the pattern of deformation recorded after the end of the eruption strongly suggests a significant contribution of the magma overpressure generated by the gas boiling, thus outlining the importance of volatiles content in magma.