Protection from Fas-mediated apoptosis by a soluble form of the Fas molecule.

Fas is an apoptosis-signaling receptor molecule on the surface of a number of cell types. Molecular cloning and nucleotide sequence analysis revealed a human Fas messenger RNA variant capable of encoding a soluble Fas molecule lacking the transmembrane domain because of the deletion of an exon encoding this region. The expression of soluble Fas was confirmed by flow cytometry and immunocytochemical analysis. Supernatants from cells transfected with the variant messenger RNA blocked apoptosis induced by the antibody to Fas. Levels of soluble Fas were elevated in patients with systemic lupus erythematosus, and mice injected with soluble Fas displayed autoimmune features.

Prevention of rat neonatal cardiomyocyte apoptosis induced by simulated in vitro ischemia and reperfusion.

Apoptosis, or programmed cell death, is an active metabolic response to physiological signals or exposure to cytotoxic agents. Recent evidence has shown that the cell death response can be modified by agents presumed to be unrelated to the initial signal, but capable of interfering with the molecular mechanisms of the apoptotic pathway progression. Here we show the results of investigations on the use of a phospholipid-based pharmaceutical preparation for suppression of myocardial damage. First, we show that serum or serum/glucose deprivation, in vitro ischemia with subsequent simulated reperfusion, inhibition of protein synthesis, and treatment with ceramide, staurosporine, adriamycin, cis-platinum and menadione induce apoptotic death in a primary culture of rat neonatal cardiomyocytes. Then we demonstrate that a mixture of specific phospholipids, which has been originally purified from soy flour on the basis of its anti-apoptotic activity, prevents cardiomyocyte death induced by serum or serum/glucose deprivation, by ischemia with subsequent simulated reperfusion, and by ceramide, but not by other cytotoxic treatments. This suggests that ceramide, a lipid secondary messenger which triggers apoptosis induced by some cytotoxic agents, may be involved in the process of signaling ischemia/reperfusion induced apoptotic death of cardiomyocytes. These results further demonstrate that an active pharmaceutical preparation for the suppression of cardiomyocyte death can be formulated based upon a novel strategy of apoptosis modification.

Dehydrothalebanin: a source of resistance from Glycosmis pentaphylla against the citrus root weevil Diaprepes abbreviatus.

Roots of a citrus relative, Glycosmis pentaphylla (orangeberry), were shown to inhibit the growth and survival of larvae of the citrus root weevil Diaprepes abbreviatus. Roots of G. pentaphyllaincorporated into the diet of D. abbreviatus increasingly inhibited the growth of neonate larvae with increased concentration of roots, while roots from citrus rootstocks produced little inhibition. The diet-incorporation assay was used to guide fractionation of an active acetone extract of G.pentaphylla roots. Three major fractions from silica open-column liquid chromatography were active, and these were purified using semipreparative normal-phase HPLC. A single active HPLC subfraction was isolated from each of the three liquid chromatography fractions, and two active compounds were isolated and identified by GC-MSD. GC-MSD and NMR identified one compound as the amide dehydrothalebanin B, and the other was identified by GC-MSD as dieldrin, a chlorinated hydrocarbon insecticide whose origin in our samples is uncertain.

The impact of maternal history and behavior upon foster placement and adjustment in sexually abused girls.

This study investigated the impact of the nonoffending mother's childhood history and current functioning upon the psychological status and placement decisions for 68 sexually abused girls. Maternal history of abuse and/or poor childhood attachment relationships were significantly related to current maternal substance abuse. Maternal substance abuse and dissatisfaction with social support were significantly associated with lack of maternal support to the child and more abuse incidents, which in turn were related to more sexual abuse-related symptomatology and placement in foster care. The findings are congruent with studies indicating that intergenerational transmission of abuse is not inevitable, and suggest that maternal substance abuse and social isolation are important mediating variables between maternal history of sexual abuse and response to the abused child. The findings suggest that interventions targeted to the nonoffending mothers may be important in reducing child dysfunction and placement in foster care in the aftermath of sexual abuse.

Identification of sources of plant resistance to Diaprepes abbreviatus (Coleoptera: Curculionidae) by three bioassays.

Host plant resistance to the root weevil Diaprepes abbreviatus (L.) was assessed for 3 citrus rootstock cultivars, 5 promising hybrid rootstocks, and 3 citroid fruit trees using 3 bioassay methods: a pot bioassay with 1-yr seedlings; a new, 21-cm plastic cell bioassay with 5-mo seedlings; and a diet incorporation bioassay. The plastic cell bioassay is a more rapid screening method and is capable of evaluating a larger number of entries in a shorter period compared with current methods. The 3 bioassays yielded similar results. Larval growth was inhibited by 2 of the remote citroid fruit trees, Murraya koenigii (L.) Sprengel and Glycosmis pentaphylla (Retzius) Correa, compared with growth on commercial rootstock cultivars. Specifically, larvae allowed to feed on roots of M. koenigii or G. pentaphylla gained less weight compared with larvae fed on the commercial rootstock cultivar 'Swingle' [Citrus paradisi Macfayden x Poncirus trifoliata (L.) Rafinesque-Schmaltz]. The resistance of G. pentaphylla confirms previous reports. M. koenigii is a new source of resistance to D. abbreviatus.

Long-acting progestin-only contraceptives enhance human endometrial stromal cell expressed neuronal pentraxin-1 and reactive oxygen species to promote endothelial cell apoptosis.

CONTEXT: Despite the absence of progesterone receptor protein in human endometrial endothelial cells (HEECs), endometria of women receiving long-acting progestin-only contraceptives (LAPCs) display reduced uterine blood flow, elevated reactive oxygen species generation, increased angiogenesis, and irregularly distributed, enlarged, fragile microvessels resulting in abnormal uterine bleeding. OBJECTIVE: We propose that paracrine factors from LAPC-treated human endometrial stromal cells (HESCs) impair HEEC functions by shifting the balance between HEEC viability and death in favor of the latter. DESIGN AND SETTING: Proliferation, apoptosis, and transcriptome analyses were performed in HEECs treated with conditioned medium supernatant (CMS) derived from HESCs treated with estradiol (E2) ± medroxyprogesterone acetate or etonogestrel under normoxia or hypoxia. Mass spectrometry interrogated the CMS secretome while immunostaining for neuronal pentraxin-1 (NPTX1), cleaved caspase-3, and cytochrome c was performed in cultured HEECs and paired endometria from women using LAPCs. MAIN OUTCOME: HEEC apoptosis and its underlying mechanism. RESULTS: HESC CMS from E2 + medroxyprogesterone acetate or E2 + etonogestrel incubations under hypoxia induced HEEC apoptosis (P < .05), whereas mass spectrometry of the CMS revealed increased NPTX1 secretion (P < .05). Endothelial cleaved caspase-3 and stromal NPTX1 immunoreactivity were significantly higher in LAPC-treated endometria (P < .001). Transcriptomics revealed AKT signaling inhibition and mitochondrial dysfunction in HEECs incubated with HESC CMS. In vitro analyses proved that CMS decreased HEEC AKT phosphorylation (P < .05) and that recombinant NPTX1 (P < .05) or NPTX1 + H2O2 (P < .001) increase HEEC apoptosis and cytosolic cytochrome c levels. CONCLUSIONS: LAPC-enhanced NPTX1 secretion and reactive oxygen species generation in HESCs impair HEEC survival resulting in a loss in vascular integrity, demonstrating a novel paracrine mechanism to explain LAPC-induced abnormal uterine bleeding.