Binding of (chloroethyl 3H) cyclophosphamide to rat embryo proteins: two-dimensional gel analysis.

Cells of the heart of the day 10 rat embryo are resistant to the cytotoxic effects of cyclophosphamide (CP) while cells of neuroepithelium (head) are sensitive. Using radiolabeled CP we have investigated the hypothesis that this difference in sensitivity is related to the binding (alkylation) of drug to specific proteins in heart and head cells. Two-dimensional gel analysis reveals that CP alkylates a variety of proteins in both heart and head cells. Although minor differences in the profiles of alkylated proteins exist between heart and head cells, the major proteins alkylated by CP in the two populations are identical. These data, therefore, do not support the hypothesis that sensitivity to the cytotoxic effects of CP is related to drug binding to specific proteins.     

Mitochondrial Staining Allows Robust Elimination of Apoptotic and Damaged Cells during Cell Sorting

High-speed fluorescence-activated cell sorting is relevant for a plethora of applications, such as PCR-based techniques, microarrays, cloning, and propagation of selected cell populations. We suggest a simple cell-sorting technique to eliminate early and late apoptotic and necrotic cells, with good signal-to-noise ratio and a high-purity yield. The mitochondrial potential dye, TMRE (tetramethylrhodamine ethyl ester perchlorate), was used to separate viable and non-apoptotic cells from the cell sorting samples. TMRE staining is reversible and does not affect cell proliferation and viability. Sorted TMRE⁺ cells contained a negligible percentage of apoptotic and damaged cells and had a higher proliferative potential as compared with their counterpart cells, sorted on the basis of staining with DNA viability dye. This novel sorting technique using TMRE does not interfere with subsequent functional assays and is a method of choice for the enrichment of functionally active, unbiased cell populations.     

Agr2‐interacting Prod1‐like protein Tfp4 from Xenopus laevis is necessary for early forebrain and eye development as well as for the tadpole appendage regeneration

The Agr family genes, Ag1, Agr2, and Agr3, encode for the thioredoxin domain containing secreted proteins and are specific only for vertebrates. These proteins are attracting increasing attention due to their involvement in many physiological and pathological processes, including exocrine secretion, cancer, regeneration of the body appendages, and the early brain development. At the same time, the mode by which Agrs regulate intracellular processes are poorly understood. Despite that the receptor to Agr2, the membrane anchored protein Prod1, has been firstly discovered in Urodeles, and it has been shown to interact with Agr2 in the regenerating limb, no functional homologs of Prod1 were identified in other vertebrates. This raises the question of the mechanisms by which Agrs can regulate regeneration in other lower vertebrates. Recently, we have identified that Tfp4 (three‐fingers Protein 4), the structural and functional homolog of Prod1 in Anurans, interacts with Agr2 in Xenopus laevis embryos. In the present work we show by several methods that the activity of Tfp4 is essential for the tadpole tail regeneration as well as for the early eye and forebrain development during embryogenesis. These data show for the first time the common molecular mechanism of regeneration regulation in amphibians by interaction of Prod1 and Agr2 proteins.     

Relationship between suicide and myocardial infarction with regard to changing physical environmental conditions

In recent years, the possible association of changes in mortality from cardiovascular disease and myocardial infarction (MI) and deaths related to violence and the suicide rate has been repeatedly discussed. This study examined the relationship between cosmic physical changes (solar, geomagnetic and other space activity parameters) and changes in the total number of in-hospital and MI-related deaths and deaths from suicide to determine if a relationship exists between the distribution of total and MI-related deaths with suicide over time; some differences in the serotonergic mechanisms involved in the pathogenesis of MI and suicide were also taken into account. All suicides (n=2359) registered in the State of Israel from 1981 to 1989 (108 months) were analysed and compared with the total number of deaths (n=15601) and deaths from MI (n=1573) in a large university hospital over 180 months (1974–1989). The following were the main features of the Results. (1) Monthly suicide rate was correlated with space proton flux (r=0.42,P=0.0001) and with geomagnetic activity (r=–0.22,P=0.03). (2) Total hospital and MI-related deaths were correlated with solar activity parameters (r=0.35,P<0.001) and radiowave propagation (r=0.52-0.44,P<0.001), an with proton flux (r=–0.3 to –0.26,P<0.01). (3) Monthly suicide distribution over 108 months was correlated with MI (r=–0.33,P=0.0005) and total hospital mortality (r=–0.22,P=0.024). (4) Gender differences were prominent. We conclude that the monthly distributions of suicides and deaths from MI are adversely related to many environmental physical parameters and negatively correlated with each other.