High-resolution measurement of breaks in prematurely condensed chromosomes by differential staining

A relatively simple method has been developed to improve the resolution for measuring breaks produced in interphase chromosomes by X rays or other agents following the induction of premature chromosome condensation (PCC). Mitotic HeLa cells, which induce PCC when fused with interphase cells, were obtained from cultures grown for several generations in 5-bromodeoxyuridine (BrdU). These were fused to cells from low-passage confluent cultures of normal human fibroblasts and subsequently stained by a modified fluorescence-plus-Giemsa (FPG) technique. Following this protocol the prematurely condensed chromosomes stain intensely, whereas the mitotic chromosomes of the inducer cell(s), which are intermingled with them, stain very lightly. With this technique the interphase chromosomes and their fragments can be identified unequivocally, making scoring much easier and more accurate. The frequency of breaks produced in G1 phase AG1522 human fibroblasts immediately following X-ray doses of 58 and 117 rad was 3.68 and 7.38 per cell, respectively. Use of this technique should allow the detection of damage from ionizing radiation at doses lower than 10 rad.     

Epididymal storage at abdominal temperature reduces the time required for capacitation of hamster spermatozoa

The epididymis was reflected unilaterally or bilaterally to the abdomen in adult hamsters, leaving normally functioning testes in the scrotum. In unilateral cases, spermatozoa taken from the abdominal cauda, 1 month or more post-operatively, underwent a reversal of head agglutination and dispersed earlier, and underwent hyperactivation and fertilized cumulus-free eggs about 30-45 min sooner than did spermatozoa from the contralateral scrotal cauda. In addition, spermatozoa from the abdominal cauda began to undergo a spontaneous acrosome reaction 30-45 min earlier and to a greater extent than in control spermatozoa. Finally, in females mated at or soon after ovulation, spermatozoa ejaculated by bilaterally cryptepididymal males fertilized eggs 30-45 min before those from normal males. Other females mated to bilaterally cryptepididymal males gave birth to normal litters. The results are considered in terms of the possibility that temperature-sensitive sperm-binding macromolecules, which may be involved in sperm storage in the cauda epididymis, could be one determinant of the need for capacitation.     

Caspase 3-mediated stimulation of tumor cell repopulation during cancer radiotherapy

In cancer treatment, apoptosis is a well-recognized cell death mechanism through which cytotoxic agents kill tumor cells. Here we report that dying tumor cells use the apoptotic process to generate potent growth-stimulating signals to stimulate the repopulation of tumors undergoing radiotherapy. Furthermore, activated caspase 3, a key executioner in apoptosis, is involved in the growth stimulation. One downstream effector that caspase 3 regulates is prostaglandin E(2) (PGE(2)), which can potently stimulate growth of surviving tumor cells. Deficiency of caspase 3 either in tumor cells or in tumor stroma caused substantial tumor sensitivity to radiotherapy in xenograft or mouse tumors. In human subjects with cancer, higher amounts of activated caspase 3 in tumor tissues are correlated with markedly increased rate of recurrence and death. We propose the existence of a cell death-induced tumor repopulation pathway in which caspase 3 has a major role.